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Chapter 17

From Gene to Protein

PowerPoint Lectures for
Biology, Seventh Edition
Neil Campbell and Jane Reece

Lectures by Chris Romero
Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
• Overview: The Flow of Genetic Information
• The information content of DNA
– Is in the form of specific sequences of
nucleotides along the DNA strands

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• The DNA inherited by an organism
– Leads to specific traits by dictating the
synthesis of proteins

• The process by which DNA directs protein
synthesis, gene expression
– Includes two stages, called transcription and
translation

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• The ribosome
– Is part of the cellular machinery for translation,
polypeptide synthesis

Figure 17.1
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• Concept 17.1: Genes specify proteins via
transcription and translation

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Evidence from the Study of Metabolic Defects
• In 1909, British physician Archibald Garrod
– Was the first to suggest that genes dictate
phenotypes through enzymes that catalyze
specific chemical reactions in the cell

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Nutritional Mutants in Neurospora: Scientific Inquiry

• Beadle and Tatum causes bread mold to
mutate with X-rays
– Creating mutants that could not survive on
minimal medium

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• Using genetic crosses
– They determined that their mutants fell into three
classes, each mutated in a different gene
EXPERIMENT

RESULTS

Working with the mold Neurospora crassa, George Beadle and Edward Tatum had isolated mutants requiring
arginine in their growth medium and had shown genetically that these mutants fell into three classes, each
defective in a different gene. From other considerations, they suspected that the metabolic pathway of arginine
biosynthesis included the precursors ornithine and citrulline. Their most famous experiment, shown here,
tested both their one gene–one enzyme hypothesis and their postulated arginine pathway. In this experiment,
they grew their three classes of mutants under the four different conditions shown in the Results section below.
The wild-type strain required only the minimal medium for growth. The three classes of mutants had
different growth requirements
Wild type
Minimal
medium
(MM)
(control)
MM +
Ornithine
MM +
Citrulline

Figure 17.2

MM +
Arginine
(control)

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Class I
Mutants

Class II
Mutants

Class III
Mutants
CONCLUSION

From the growth patterns of the mutants, Beadle and Tatum deduced that each mutant was unable
to carry out one step in the pathway for synthesizing arginine, presumably because it lacked the
necessary enzyme. Because each of their mutants was mutated in a single gene, they concluded that
each mutated gene must normally dictate the production of one enzyme. Their results supported the
one gene–one enzyme hypothesis and also confirmed the arginine pathway.
(Notice that a mutant can grow only if supplied with a compound made after the defective step.)

Wild type
Precursor
Gene A

Class I
Mutants
(mutation
in gene A)
Precursor

Precursor

Precursor

A

A

A

Ornithine

Ornithine

Ornithine

B

B

B

Citrulline

Citrulline

Citrulline

C

C

C

Arginine

Arginine

Arginine

Enzyme
A
Ornithine

Gene B

Enzyme
B
Citrulline

Gene C

Enzyme
C
Arginine

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Class II
Mutants
(mutation
in gene B)

Class III
Mutants
(mutation
in gene C)
• Beadle and Tatum developed the “one gene–
one enzyme hypothesis”
– Which states that the function of a gene is to
dictate the production of a specific enzyme

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The Products of Gene Expression: A Developing Story

• As researchers learned more about proteins
– The made minor revision to the one gene–one
enzyme hypothesis

• Genes code for polypeptide chains or for RNA
molecules

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Basic Principles of Transcription and Translation
• Transcription
– Is the synthesis of RNA under the direction of
DNA
– Produces messenger RNA (mRNA)

• Translation
– Is the actual synthesis of a polypeptide, which
occurs under the direction of mRNA
– Occurs on ribosomes

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• In prokaryotes
– Transcription and translation occur together

TRANSCRIPTION

DNA
mRNA
Ribosome

TRANSLATION
Polypeptide

(a) Prokaryotic cell. In a cell lacking a nucleus, mRNA
produced by transcription is immediately translated
without additional processing.

Figure 17.3a
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• In eukaryotes
– RNA transcripts are modified before becoming
true mRNA
Nuclear
envelope

DNA

TRANSCRIPTION

Pre-mRNA

RNA PROCESSING

mRNA

Ribosome
TRANSLATION
Polypeptide

Figure 17.3b
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(b) Eukaryotic cell. The nucleus provides a separate
compartment for transcription. The original RNA
transcript, called pre-mRNA, is processed in various
ways before leaving the nucleus as mRNA.
• Cells are governed by a cellular chain of
command
– DNA → RNA → protein

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The Genetic Code
• How many bases correspond to an amino
acid?

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Codons: Triplets of Bases
• Genetic information
– Is encoded as a sequence of nonoverlapping
base triplets, or codons

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• During transcription
– The gene determines the sequence of bases
along the length of an mRNA molecule
Gene 2

DNA
molecule

Gene 1
Gene 3

DNA strand 3′
5′
A C C A A A C C G A G T
(template)
TRANSCRIPTION

mRNA

5′

U G G U U U G G C U C A
Codon

TRANSLATION

Protein

Figure 17.4

Trp
Amino acid

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Phe

Gly

Ser

3′
Cracking the Code
• A codon in messenger RNA

Figure 17.5

Second mRNA base
U
C
A
UAU
UUU
UCU
Tyr
Phe
UAC
UUC
UCC
U
UUA
UCA Ser UAA Stop
UAG Stop
UUG Leu UCG
CUU
CUC
C
CUA
CUG

CCU
CCC
Leu CCA
CCG

Pro

AUU
AUC
A
AUA
AUG

ACU
ACC
ACA
ACG

Thr

GUU
G GUC
GUA
GUG

lle
Met or
start

GCU
GCC
Val
GCA
GCG

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Ala

G

U
UGU
Cys
UGC
C
UGA Stop A
UGG Trp G
U
CAU
CGU
His
CAC
CGC
C
Arg
CAA
CGA
A
Gln
CAG
CGG
G
U
AAU
AGU
Asn
AAC
AGC Ser C
A
AAA
AGA
Lys
AAG
AGG Arg G
U
GAU
GGU
C
GAC Asp GGC
Gly
GAA
GGA
A
Glu GGG
GAG
G

Third mRNA base (3′ end)

First mRNA base (5′ end)

– Is either translated into an amino acid or serves as
a translational stop signal
• Codons must be read in the correct reading
frame
– For the specified polypeptide to be produced

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Evolution of the Genetic Code
• The genetic code is nearly universal
– Shared by organisms from the simplest
bacteria to the most complex animals

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• In laboratory experiments
– Genes can be transcribed and translated after
being transplanted from one species to
another

Figure 17.6
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• Concept 17.2: Transcription is the DNAdirected synthesis of RNA: a closer look

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Molecular Components of Transcription
• RNA synthesis
– Is catalyzed by RNA polymerase, which pries
the DNA strands apart and hooks together the
RNA nucleotides
– Follows the same base-pairing rules as DNA,
except that in RNA, uracil substitutes for
thymine

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Synthesis of an RNA Transcript
• The stages of transcription are
Promoter

– Initiation

Transcription unit

5′
3′

3′
5′

Start point
RNA polymerase

– Elongation
– Termination

DNA
Initiation. After RNA polymerase binds to
the promoter, the DNA strands unwind, and
the polymerase initiates RNA synthesis at the
start point on the template strand.

1

5′
3′

Unwound
DNA

3′
5′

Template strand of
DNA
transcript
2 Elongation. The polymerase moves downstream, unwinding the
DNA and elongating the RNA transcript 5′ → 3 ′. In the wake of
transcription, the DNA strands re-form a double helix.
Rewound
RNA

RNA
5′
3′

3′
5′

3′
5′

RNA
transcript

3 Termination. Eventually, the RNA
transcript is released, and the
polymerase detaches from the DNA.

5′
3′

3′
5′
5′

Figure 17.7
Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings

Completed RNA
transcript

3′
Non-template
strand of DNA

Elongation

RNA nucleotides
RNA
polymerase

A
3′

T

C

C

C

A

A

T

U

3′ end
G

T

A

U
C

A
T

E
A

G
G

C

A

G

T

A

A

T

Direction of transcription
(“downstream”)

5′

Newly made
RNA

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G

5′

T

G

C

A

Template
strand of DNA
RNA Polymerase Binding and Initiation of Transcription

• Promoters signal the initiation of RNA synthesis
• Transcription factors
– Help eukaryotic RNA polymerase recognize
promoter sequences
1 Eukaryotic promoters

TRANSCRIPTION

DNA

RNA PROCESSING

Pre-mRNA

mRNA
TRANSLATION

Ribosome
Polypeptide

5′
3′

Promoter

3′
5′

T A T A A AA
AT A T T T T

TATA box

Start point
2

Template
DNA strand
Several transcription
factors

Transcription
factors

5′
3′

3 Additional transcription

3′
5′

factors

RNA polymerase II

5′
3′

Figure 17.8
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Transcription factors

3′
5′

5′

RNA transcript
Transcription initiation complex
Elongation of the RNA Strand
• As RNA polymerase moves along the DNA
– It continues to untwist the double helix,
exposing about 10 to 20 DNA bases at a time
for pairing with RNA nucleotides

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Termination of Transcription
• The mechanisms of termination
– Are different in prokaryotes and eukaryotes

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• Concept 17.3: Eukaryotic cells modify RNA
after transcription
• Enzymes in the eukaryotic nucleus
– Modify pre-mRNA in specific ways before the
genetic messages are dispatched to the
cytoplasm

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Alteration of mRNA Ends
• Each end of a pre-mRNA molecule is modified
in a particular way
– The 5′ end receives a modified nucleotide cap
– The 3′ end gets a poly-A tail

A modified guanine nucleotide
added to the 5′ end
TRANSCRIPTION

RNA PROCESSING

50 to 250 adenine nucleotides
added to the 3′ end

DNA
Pre-mRNA

mRNA

5′

Protein-coding segment

3′

G P P P

AAUAAA

Ribosome
TRANSLATION

5′ Cap
Polypeptide

Polyadenylation signal

5′ UTR

Start codon Stop codon

Figure 17.9
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3′ UTR

AAA…AAA
Poly-A tail
Split Genes and RNA Splicing
• RNA splicing
– Removes introns and joins exons

TRANSCRIPTION

RNA PROCESSING

DNA
Pre-mRNA

5′ Exon Intron
Pre-mRNA 5′ Cap
30
31
1

Coding
segment

mRNA
Ribosome

Intron

Exon

Exon

3′
Poly-A tail

104

105

146

Introns cut out and
exons spliced together

TRANSLATION

Polypeptide

mRNA 5′ Cap

1
3′ UTR

Figure 17.10
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Poly-A tail
146

3′ UTR
• Is carried out by spliceosomes in some cases
5′

1

RNA transcript (pre-mRNA)
Intron

Exon 1

Exon 2

Protein
Other proteins

snRNA
snRNPs

Spliceosome

2

5′

Spliceosome
components

3

Figure 17.11

5′

mRNA
Exon 1

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Exon 2

Cut-out
intron
Ribozymes
• Ribozymes
– Are catalytic RNA molecules that function as
enzymes and can splice RNA

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The Functional and Evolutionary Importance of Introns

• The presence of introns
– Allows for alternative RNA splicing

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• Proteins often have a modular architecture
– Consisting of discrete structural and functional
regions called domains

• In many cases
– Different exons code for the different domains in a
protein
Gene
DNA

Exon 1 Intron Exon 2
Transcription
RNA processing

Intron Exon 3

Translation
Domain 3
Domain 2
Domain 1

Figure 17.12
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Polypeptide
• Concept 17.4: Translation is the RNA-directed
synthesis of a polypeptide: a closer look

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Molecular Components of Translation
• A cell translates an mRNA message into
protein
– With the help of transfer RNA (tRNA)

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• Translation: the basic concept
TRANSCRIPTION

DNA
mRNA
Ribosome

TRANSLATION
Polypeptide

Amino
acids

Polypeptide

Trp
Ph e

tRNA with
amino acid
Ribosome attached
Gly
tRNA

A

C

GC

C

A A A
U G G U U U G G C

Codons

5′
Figure 17.13

mRNA

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G

Anticodon

3′
• Molecules of tRNA are not all identical
– Each carries a specific amino acid on one end
– Each has an anticodon on the other end

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The Structure and Function of Transfer RNA
• A tRNA molecule
A
– Consists of a single RNA strand C
that is only
C
about 80 nucleotides long

– Is roughly L-shaped

3′

A
C
C
A 5′
C G
G C
C G
U G
U A
A U
A U
U C
UA
C A C AG
*
G
*
G U G U *
C
C
* *
U C
*
* G AG C
(a) Two-dimensional structure. The four base-paired regions and
G C
U A
three loops are characteristic of all tRNAs, as is the base sequence
* G
of the amino acid attachment site at the 3′ end. The anticodon triplet
A
A*
is unique to each tRNA type. (The asterisks mark bases that have
C
U
*
been chemically modified, a characteristic of tRNA.)
A
G
A

Figure 17.14a
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Amino acid
attachment site

Anticodon

C U C
G A G

A G *
*
G
A G G

Hydrogen
bonds
5′
3′

Amino acid
attachment site

Hydrogen
bonds

A AG
3′
Anticodon
(b) Three-dimensional structure

Figure 17.14b
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5′

Anticodon

(c) Symbol used
in this book
• A specific enzyme called an aminoacyl-tRNA
synthetase
– Joins each amino acid to the correct tRNA
Amino acid

P P

Aminoacyl-tRNA
synthetase (enzyme)
1 Active site binds the
amino acid and ATP.

P Adenosine

ATP

2 ATP loses two P groups
and joins amino acid as AMP.
P

Pyrophosphate
Pi

Phosphates
3 Appropriate
tRNA covalently
Bonds to amino
Acid, displacing
AMP.

P

Adenosine

Pi

Pi

tRNA

P Adenosine

AMP

4 Activated amino acid
is released by the enzyme.

Figure 17.15

Aminoacyl tRNA
(an “activated
amino acid”)

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Ribosomes
• Ribosomes
– Facilitate the specific coupling of tRNA
anticodons with mRNA codons during protein
synthesis

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• The ribosomal subunits
– Are constructed of proteins and RNA
molecules named ribosomal RNA or rRNA
DNA

TRANSCRIPTION

mRNA
Ribosome
TRANSLATION

Polypeptide

Growing
polypeptide

Exit tunnel

tRNA
molecules

Large
subunit
E

P A
Small
subunit

5′
mRNA

Figure 17.16a

3′

(a) Computer model of functioning ribosome. This is a model of a bacterial
ribosome, showing its overall shape. The eukaryotic ribosome is roughly
similar. A ribosomal subunit is an aggregate of ribosomal RNA molecules
and proteins.

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• The ribosome has three binding sites for tRNA
– The P site
– The A site
– The E site

P site (Peptidyl-tRNA
binding site)
A site (AminoacyltRNA binding site)

E site
(Exit site)

E

mRNA
binding site

Figure 17.16b

P

A

Large
subunit

Small
subunit

(b) Schematic model showing binding sites. A ribosome has an mRNA
binding site and three tRNA binding sites, known as the A, P, and E
sites. This schematic ribosome will appear in later diagrams.

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Amino end

Growing polypeptide
Next amino acid
to be added to
polypeptide chain

tRNA
3′

mRNA

5′

Codons

(c) Schematic model with mRNA and tRNA. A tRNA fits into a binding site when its anticodon
base-pairs with an mRNA codon. The P site holds the tRNA attached to the growing
polypeptide. The A site holds the tRNA carrying the next amino acid to be added to the
polypeptide chain. Discharged tRNA leaves via the E site.
Figure 17.16c
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Building a Polypeptide
• We can divide translation into three stages
– Initiation
– Elongation
– Termination

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Ribosome Association and Initiation of Translation
• The initiation stage of translation
– Brings together mRNA, tRNA bearing the first
amino acid of the polypeptide, and two
subunits of a ribosome
t
Me

P site

3′ U A C 5′
5′ A U G 3′

Initiator tRNA

t
Me

GTP

GDP

E

mRNA
5′

Start codon

mRNA binding site

Figure 17.17

3′
Small
ribosomal
subunit

1 A small ribosomal subunit binds to a molecule of
mRNA. In a prokaryotic cell, the mRNA binding site
on this subunit recognizes a specific nucleotide
sequence on the mRNA just upstream of the start
codon. An initiator tRNA, with the anticodon UAC,
base-pairs with the start codon, AUG. This tRNA
carries the amino acid methionine (Met).

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Large
ribosomal
subunit

5′

A
3′

Translation initiation complex

2 The arrival of a large ribosomal subunit completes
the initiation complex. Proteins called initiation
factors (not shown) are required to bring all the
translation components together. GTP provides
the energy for the assembly. The initiator tRNA is
in the P site; the A site is available to the tRNA
bearing the next amino acid.
Elongation of the Polypeptide Chain
• In the elongation stage of translation
– Amino acids are added one by one to the
preceding amino acid
TRANSCRIPTION

Amino end
of polypeptide

DNA
mRNA
Ribosome

TRANSLATION

Polypeptide

mRNA
Ribosome ready for
next aminoacyl tRNA

E
3′
P A
site site

5′

1 Codon recognition. The anticodon
of an incoming aminoacyl tRNA
base-pairs with the complementary
mRNA codon in the A site. Hydrolysis
of GTP increases the accuracy and
efficiency of this step.

2 GTP
2 GDP

E

E
P

Figure 17.18

3 Translocation. The ribosome
translocates the tRNA in the A
site to the P site. The empty tRNA
in the P site is moved to the E site,
where it is released. The mRNA
moves along with its bound tRNAs,
bringing the next codon to be
translated into the A site.

P

A
GDP
GTP

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E
P

A

A
2 Peptide bond formation. An
rRNA molecule of the large
subunit catalyzes the formation
of a peptide bond between the
new amino acid in the A site and
the carboxyl end of the growing
polypeptide in the P site. This step
attaches the polypeptide to the
tRNA in the A site.
Termination of Translation
• The final stage of translation is termination
– When the ribosome reaches a stop codon in
the mRNA
Release
factor

Free
polypeptide
5′
3′

3′
5′

5′

3′

Stop codon
(UAG, UAA, or UGA)
1 When a ribosome reaches a stop 2 The release factor hydrolyzes 3 The two ribosomal subunits
codon on mRNA, the A site of the the bond between the tRNA in and the other components of
ribosome accepts a protein called the P site and the last amino
the assembly dissociate.
a release factor instead of tRNA.
acid of the polypeptide chain.
The polypeptide is thus freed
from the ribosome.
Figure 17.19
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Polyribosomes
• A number of ribosomes can translate a single
mRNA molecule simultaneously
– Forming a polyribosome

Completed
polypeptide

Growing
polypeptides
Incoming
ribosomal
subunits
Start of
mRNA
(5′ end)

Polyribosom
e

End of
mRNA
(3′ end)
(a) An mRNA molecule is generally translated simultaneously
by several ribosomes in clusters called polyribosomes.

Ribosomes
mRNA

0.1 µm

Figure 17.20a, b

(b) This micrograph shows a large polyribosome in a prokaryotic
cell (TEM).

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Completing and Targeting the Functional Protein
• Polypeptide chains
– Undergo modifications after the translation
process

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Protein Folding and Post-Translational Modifications

• After translation
– Proteins may be modified in ways that affect
their three-dimensional shape

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Targeting Polypeptides to Specific Locations
• Two populations of ribosomes are evident in
cells
– Free and bound

• Free ribosomes in the cytosol
– Initiate the synthesis of all proteins

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• Proteins destined for the endomembrane
system or for secretion
– Must be transported into the ER
– Have signal peptides to which a signalrecognition particle (SRP) binds, enabling the
translation ribosome to bind to the ER

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• The signal mechanism for targeting proteins to
the ER
1 Polypeptide
synthesis begins
on a free
ribosome in
the cytosol.

2 An SRP binds
to the signal
peptide, halting
synthesis
momentarily.

3 The SRP binds to a
receptor protein in the ER
membrane. This receptor
is part of a protein complex
(a translocation complex)
that has a membrane pore
and a signal-cleaving enzyme.

4 The SRP leaves, and
the polypeptide resumes
growing, meanwhile
translocating across the
membrane. (The signal
peptide stays attached
to the membrane.)

5 The signalcleaving
enzyme
cuts off the
signal peptide.

6 The rest of
the completed
polypeptide leaves
the ribosome and
folds into its final
conformation.

Ribosome

mRNA
Signal
peptide
Signalrecognition
particle
(SRP) SRP
receptor
CYTOSOL protein

ERLUMEN

Translocation
complex

Figure 17.21
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Signal
peptide
removed

ER
membrane
Protein
• Concept 17.5: RNA plays multiple roles in the
cell: a review
• RNA
– Can hydrogen-bond to other nucleic acid
molecules
– Can assume a specific three-dimensional
shape
– Has functional groups that allow it to act as a
catalyst
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• Types of RNA in a Eukaryotic Cell

Table 17.1
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• Concept 17.6: Comparing gene expression in
prokaryotes and eukaryotes reveals key differences
• Prokaryotic cells lack a nuclear envelope
– Allowing translation to begin while transcription is
still in progress
RNA polymerase
DNA

mRNA

Polyribosome
RNA
polymerase

Direction of
transcription

DNA

Polyribosome
Polypeptide
(amino end)
Ribosome

Figure 17.22

0.25 µm

mRNA (5′ end)

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• In a eukaryotic cell
– The nuclear envelope separates transcription
from translation
– Extensive RNA processing occurs in the
nucleus

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• Concept 17.7: Point mutations can affect
protein structure and function
• Mutations
– Are changes in the genetic material of a cell

• Point mutations
– Are changes in just one base pair of a gene

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• The change of a single nucleotide in the DNA’s
template strand
– Leads to the production of an abnormal protein
Wild-type hemoglobin DNA
3′

Mutant hemoglobin DNA

C T

T

G U A

5′

In the DNA, the
mutant template
strand has an A where
the wild-type template
has a T.

The mutant mRNA has
a U instead of an A in
one codon.

3′

5′

T

C A

mRNA

mRNA
G A

A

5′

3′

5′

3′

Normal hemoglobin

Sickle-cell hemoglobin

Glu

Val

Figure 17.23
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The mutant (sickle-cell)
hemoglobin has a valine
(Val) instead of a glutamic
acid (Glu).
Types of Point Mutations
• Point mutations within a gene can be divided
into two general categories
– Base-pair substitutions
– Base-pair insertions or deletions

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Substitutions
• A base-pair substitution
– Is the replacement of one nucleotide and its
partner with another pair of nucleotides
– Can cause missense or nonsense
Wild type
mRNA
Protein

5′

A U G
Met

A A G U U U GG C U A A
Lys

Phe

Gly

3′

Stop

Amino end

Carboxyl end
Base-pair substitution
No effect on amino acid sequence
U instead of C
A U G A A G U U U G G U U A A
Met

Lys

Missense

Phe

Gly

Stop

A instead of G

A U G A A G U U U A G U U A A
Met

Lys

Phe

Ser

Stop

Nonsense
U instead of A
A U G U A G U U U G G C U A A

Figure 17.24

Met

Stop

Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
Insertions and Deletions
• Insertions and deletions
– Are additions or losses of nucleotide pairs in a
gene
– May produce frameshift mutations
Wild type

mRNA
Protein

5′

A UG A A GU U U G G C U A A
Met

Lys

Gly

Phe

Stop

Amino end
Carboxyl end
Base-pair insertion or deletion
Frameshift causing immediate nonsense

Extra U
AU G U A AG U U U G GC U A
Met

Stop

Frameshift causing
extensive missense

U Missing

A U G A A GU U G G C U A A
Met

Lys

Leu

Ala

Insertion or deletion of 3 nucleotides:
no frameshift but extra or missing amino acid

A AG

Missing

A U G U U U G G C U A A

Figure 17.25

Met

Phe

Gly

Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings

Stop

3′
Mutagens
• Spontaneous mutations
– Can occur during DNA replication,
recombination, or repair

Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
• Mutagens
– Are physical or chemical agents that can
cause mutations

Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
What is a gene? revisiting the question
• A gene
– Is a region of DNA whose final product is either
a polypeptide or an RNA molecule

Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
• A summary of transcription and translation in a
eukaryotic cell
DNA

TRANSCRIPTION

1 RNA is transcribed

from a DNA template.
3′

5′

RNA
transcript

A
lyPo

RNA
polymerase

RNA PROCESSING

Exon

2 In eukaryotes, the

RNA transcript (premRNA) is spliced and
modified to produce
mRNA, which moves
from the nucleus to the
cytoplasm.

RNA transcript
(pre-mRNA)
Intron
Aminoacyl-tRNA
synthetase

p
Ca

NUCLEUS

Amino
acid
tRNA

FORMATION OF
INITIATION COMPLEX

CYTOPLASM 3 After leaving the
nucleus, mRNA attaches
to the ribosome.

AMINO ACID ACTIVATION
4
Each amino acid
attaches to its proper tRNA
with the help of a specific
enzyme and ATP.

Growing
polypeptide

mRNA
A
lyPo

Activated
amino acid

Ribosomal
subunits

p
Ca
5′

TRANSLATION

C
AC

U
A AC

E

AAA
UG GUU U A U G
Codon

Figure 17.26
Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings

Ribosome

A succession of tRNAs
add their amino acids to
the polypeptide chain
Anticodon
as the mRNA is moved
through the ribosome
one codon at a time.
(When completed, the
polypeptide is released
from the ribosome.)
5

A
lyPo

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17 genetoprotein text

  • 1. Chapter 17 From Gene to Protein PowerPoint Lectures for Biology, Seventh Edition Neil Campbell and Jane Reece Lectures by Chris Romero Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 2. • Overview: The Flow of Genetic Information • The information content of DNA – Is in the form of specific sequences of nucleotides along the DNA strands Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 3. • The DNA inherited by an organism – Leads to specific traits by dictating the synthesis of proteins • The process by which DNA directs protein synthesis, gene expression – Includes two stages, called transcription and translation Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 4. • The ribosome – Is part of the cellular machinery for translation, polypeptide synthesis Figure 17.1 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 5. • Concept 17.1: Genes specify proteins via transcription and translation Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 6. Evidence from the Study of Metabolic Defects • In 1909, British physician Archibald Garrod – Was the first to suggest that genes dictate phenotypes through enzymes that catalyze specific chemical reactions in the cell Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 7. Nutritional Mutants in Neurospora: Scientific Inquiry • Beadle and Tatum causes bread mold to mutate with X-rays – Creating mutants that could not survive on minimal medium Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 8. • Using genetic crosses – They determined that their mutants fell into three classes, each mutated in a different gene EXPERIMENT RESULTS Working with the mold Neurospora crassa, George Beadle and Edward Tatum had isolated mutants requiring arginine in their growth medium and had shown genetically that these mutants fell into three classes, each defective in a different gene. From other considerations, they suspected that the metabolic pathway of arginine biosynthesis included the precursors ornithine and citrulline. Their most famous experiment, shown here, tested both their one gene–one enzyme hypothesis and their postulated arginine pathway. In this experiment, they grew their three classes of mutants under the four different conditions shown in the Results section below. The wild-type strain required only the minimal medium for growth. The three classes of mutants had different growth requirements Wild type Minimal medium (MM) (control) MM + Ornithine MM + Citrulline Figure 17.2 MM + Arginine (control) Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Class I Mutants Class II Mutants Class III Mutants
  • 9. CONCLUSION From the growth patterns of the mutants, Beadle and Tatum deduced that each mutant was unable to carry out one step in the pathway for synthesizing arginine, presumably because it lacked the necessary enzyme. Because each of their mutants was mutated in a single gene, they concluded that each mutated gene must normally dictate the production of one enzyme. Their results supported the one gene–one enzyme hypothesis and also confirmed the arginine pathway. (Notice that a mutant can grow only if supplied with a compound made after the defective step.) Wild type Precursor Gene A Class I Mutants (mutation in gene A) Precursor Precursor Precursor A A A Ornithine Ornithine Ornithine B B B Citrulline Citrulline Citrulline C C C Arginine Arginine Arginine Enzyme A Ornithine Gene B Enzyme B Citrulline Gene C Enzyme C Arginine Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Class II Mutants (mutation in gene B) Class III Mutants (mutation in gene C)
  • 10. • Beadle and Tatum developed the “one gene– one enzyme hypothesis” – Which states that the function of a gene is to dictate the production of a specific enzyme Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 11. The Products of Gene Expression: A Developing Story • As researchers learned more about proteins – The made minor revision to the one gene–one enzyme hypothesis • Genes code for polypeptide chains or for RNA molecules Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 12. Basic Principles of Transcription and Translation • Transcription – Is the synthesis of RNA under the direction of DNA – Produces messenger RNA (mRNA) • Translation – Is the actual synthesis of a polypeptide, which occurs under the direction of mRNA – Occurs on ribosomes Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 13. • In prokaryotes – Transcription and translation occur together TRANSCRIPTION DNA mRNA Ribosome TRANSLATION Polypeptide (a) Prokaryotic cell. In a cell lacking a nucleus, mRNA produced by transcription is immediately translated without additional processing. Figure 17.3a Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 14. • In eukaryotes – RNA transcripts are modified before becoming true mRNA Nuclear envelope DNA TRANSCRIPTION Pre-mRNA RNA PROCESSING mRNA Ribosome TRANSLATION Polypeptide Figure 17.3b Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings (b) Eukaryotic cell. The nucleus provides a separate compartment for transcription. The original RNA transcript, called pre-mRNA, is processed in various ways before leaving the nucleus as mRNA.
  • 15. • Cells are governed by a cellular chain of command – DNA → RNA → protein Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 16. The Genetic Code • How many bases correspond to an amino acid? Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 17. Codons: Triplets of Bases • Genetic information – Is encoded as a sequence of nonoverlapping base triplets, or codons Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 18. • During transcription – The gene determines the sequence of bases along the length of an mRNA molecule Gene 2 DNA molecule Gene 1 Gene 3 DNA strand 3′ 5′ A C C A A A C C G A G T (template) TRANSCRIPTION mRNA 5′ U G G U U U G G C U C A Codon TRANSLATION Protein Figure 17.4 Trp Amino acid Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Phe Gly Ser 3′
  • 19. Cracking the Code • A codon in messenger RNA Figure 17.5 Second mRNA base U C A UAU UUU UCU Tyr Phe UAC UUC UCC U UUA UCA Ser UAA Stop UAG Stop UUG Leu UCG CUU CUC C CUA CUG CCU CCC Leu CCA CCG Pro AUU AUC A AUA AUG ACU ACC ACA ACG Thr GUU G GUC GUA GUG lle Met or start GCU GCC Val GCA GCG Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Ala G U UGU Cys UGC C UGA Stop A UGG Trp G U CAU CGU His CAC CGC C Arg CAA CGA A Gln CAG CGG G U AAU AGU Asn AAC AGC Ser C A AAA AGA Lys AAG AGG Arg G U GAU GGU C GAC Asp GGC Gly GAA GGA A Glu GGG GAG G Third mRNA base (3′ end) First mRNA base (5′ end) – Is either translated into an amino acid or serves as a translational stop signal
  • 20. • Codons must be read in the correct reading frame – For the specified polypeptide to be produced Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 21. Evolution of the Genetic Code • The genetic code is nearly universal – Shared by organisms from the simplest bacteria to the most complex animals Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 22. • In laboratory experiments – Genes can be transcribed and translated after being transplanted from one species to another Figure 17.6 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 23. • Concept 17.2: Transcription is the DNAdirected synthesis of RNA: a closer look Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 24. Molecular Components of Transcription • RNA synthesis – Is catalyzed by RNA polymerase, which pries the DNA strands apart and hooks together the RNA nucleotides – Follows the same base-pairing rules as DNA, except that in RNA, uracil substitutes for thymine Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 25. Synthesis of an RNA Transcript • The stages of transcription are Promoter – Initiation Transcription unit 5′ 3′ 3′ 5′ Start point RNA polymerase – Elongation – Termination DNA Initiation. After RNA polymerase binds to the promoter, the DNA strands unwind, and the polymerase initiates RNA synthesis at the start point on the template strand. 1 5′ 3′ Unwound DNA 3′ 5′ Template strand of DNA transcript 2 Elongation. The polymerase moves downstream, unwinding the DNA and elongating the RNA transcript 5′ → 3 ′. In the wake of transcription, the DNA strands re-form a double helix. Rewound RNA RNA 5′ 3′ 3′ 5′ 3′ 5′ RNA transcript 3 Termination. Eventually, the RNA transcript is released, and the polymerase detaches from the DNA. 5′ 3′ 3′ 5′ 5′ Figure 17.7 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Completed RNA transcript 3′
  • 26. Non-template strand of DNA Elongation RNA nucleotides RNA polymerase A 3′ T C C C A A T U 3′ end G T A U C A T E A G G C A G T A A T Direction of transcription (“downstream”) 5′ Newly made RNA Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings G 5′ T G C A Template strand of DNA
  • 27. RNA Polymerase Binding and Initiation of Transcription • Promoters signal the initiation of RNA synthesis • Transcription factors – Help eukaryotic RNA polymerase recognize promoter sequences 1 Eukaryotic promoters TRANSCRIPTION DNA RNA PROCESSING Pre-mRNA mRNA TRANSLATION Ribosome Polypeptide 5′ 3′ Promoter 3′ 5′ T A T A A AA AT A T T T T TATA box Start point 2 Template DNA strand Several transcription factors Transcription factors 5′ 3′ 3 Additional transcription 3′ 5′ factors RNA polymerase II 5′ 3′ Figure 17.8 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Transcription factors 3′ 5′ 5′ RNA transcript Transcription initiation complex
  • 28. Elongation of the RNA Strand • As RNA polymerase moves along the DNA – It continues to untwist the double helix, exposing about 10 to 20 DNA bases at a time for pairing with RNA nucleotides Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 29. Termination of Transcription • The mechanisms of termination – Are different in prokaryotes and eukaryotes Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 30. • Concept 17.3: Eukaryotic cells modify RNA after transcription • Enzymes in the eukaryotic nucleus – Modify pre-mRNA in specific ways before the genetic messages are dispatched to the cytoplasm Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 31. Alteration of mRNA Ends • Each end of a pre-mRNA molecule is modified in a particular way – The 5′ end receives a modified nucleotide cap – The 3′ end gets a poly-A tail A modified guanine nucleotide added to the 5′ end TRANSCRIPTION RNA PROCESSING 50 to 250 adenine nucleotides added to the 3′ end DNA Pre-mRNA mRNA 5′ Protein-coding segment 3′ G P P P AAUAAA Ribosome TRANSLATION 5′ Cap Polypeptide Polyadenylation signal 5′ UTR Start codon Stop codon Figure 17.9 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings 3′ UTR AAA…AAA Poly-A tail
  • 32. Split Genes and RNA Splicing • RNA splicing – Removes introns and joins exons TRANSCRIPTION RNA PROCESSING DNA Pre-mRNA 5′ Exon Intron Pre-mRNA 5′ Cap 30 31 1 Coding segment mRNA Ribosome Intron Exon Exon 3′ Poly-A tail 104 105 146 Introns cut out and exons spliced together TRANSLATION Polypeptide mRNA 5′ Cap 1 3′ UTR Figure 17.10 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Poly-A tail 146 3′ UTR
  • 33. • Is carried out by spliceosomes in some cases 5′ 1 RNA transcript (pre-mRNA) Intron Exon 1 Exon 2 Protein Other proteins snRNA snRNPs Spliceosome 2 5′ Spliceosome components 3 Figure 17.11 5′ mRNA Exon 1 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Exon 2 Cut-out intron
  • 34. Ribozymes • Ribozymes – Are catalytic RNA molecules that function as enzymes and can splice RNA Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 35. The Functional and Evolutionary Importance of Introns • The presence of introns – Allows for alternative RNA splicing Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 36. • Proteins often have a modular architecture – Consisting of discrete structural and functional regions called domains • In many cases – Different exons code for the different domains in a protein Gene DNA Exon 1 Intron Exon 2 Transcription RNA processing Intron Exon 3 Translation Domain 3 Domain 2 Domain 1 Figure 17.12 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Polypeptide
  • 37. • Concept 17.4: Translation is the RNA-directed synthesis of a polypeptide: a closer look Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 38. Molecular Components of Translation • A cell translates an mRNA message into protein – With the help of transfer RNA (tRNA) Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 39. • Translation: the basic concept TRANSCRIPTION DNA mRNA Ribosome TRANSLATION Polypeptide Amino acids Polypeptide Trp Ph e tRNA with amino acid Ribosome attached Gly tRNA A C GC C A A A U G G U U U G G C Codons 5′ Figure 17.13 mRNA Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings G Anticodon 3′
  • 40. • Molecules of tRNA are not all identical – Each carries a specific amino acid on one end – Each has an anticodon on the other end Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 41. The Structure and Function of Transfer RNA • A tRNA molecule A – Consists of a single RNA strand C that is only C about 80 nucleotides long – Is roughly L-shaped 3′ A C C A 5′ C G G C C G U G U A A U A U U C UA C A C AG * G * G U G U * C C * * U C * * G AG C (a) Two-dimensional structure. The four base-paired regions and G C U A three loops are characteristic of all tRNAs, as is the base sequence * G of the amino acid attachment site at the 3′ end. The anticodon triplet A A* is unique to each tRNA type. (The asterisks mark bases that have C U * been chemically modified, a characteristic of tRNA.) A G A Figure 17.14a Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Amino acid attachment site Anticodon C U C G A G A G * * G A G G Hydrogen bonds
  • 42. 5′ 3′ Amino acid attachment site Hydrogen bonds A AG 3′ Anticodon (b) Three-dimensional structure Figure 17.14b Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings 5′ Anticodon (c) Symbol used in this book
  • 43. • A specific enzyme called an aminoacyl-tRNA synthetase – Joins each amino acid to the correct tRNA Amino acid P P Aminoacyl-tRNA synthetase (enzyme) 1 Active site binds the amino acid and ATP. P Adenosine ATP 2 ATP loses two P groups and joins amino acid as AMP. P Pyrophosphate Pi Phosphates 3 Appropriate tRNA covalently Bonds to amino Acid, displacing AMP. P Adenosine Pi Pi tRNA P Adenosine AMP 4 Activated amino acid is released by the enzyme. Figure 17.15 Aminoacyl tRNA (an “activated amino acid”) Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 44. Ribosomes • Ribosomes – Facilitate the specific coupling of tRNA anticodons with mRNA codons during protein synthesis Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 45. • The ribosomal subunits – Are constructed of proteins and RNA molecules named ribosomal RNA or rRNA DNA TRANSCRIPTION mRNA Ribosome TRANSLATION Polypeptide Growing polypeptide Exit tunnel tRNA molecules Large subunit E P A Small subunit 5′ mRNA Figure 17.16a 3′ (a) Computer model of functioning ribosome. This is a model of a bacterial ribosome, showing its overall shape. The eukaryotic ribosome is roughly similar. A ribosomal subunit is an aggregate of ribosomal RNA molecules and proteins. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 46. • The ribosome has three binding sites for tRNA – The P site – The A site – The E site P site (Peptidyl-tRNA binding site) A site (AminoacyltRNA binding site) E site (Exit site) E mRNA binding site Figure 17.16b P A Large subunit Small subunit (b) Schematic model showing binding sites. A ribosome has an mRNA binding site and three tRNA binding sites, known as the A, P, and E sites. This schematic ribosome will appear in later diagrams. Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 47. Amino end Growing polypeptide Next amino acid to be added to polypeptide chain tRNA 3′ mRNA 5′ Codons (c) Schematic model with mRNA and tRNA. A tRNA fits into a binding site when its anticodon base-pairs with an mRNA codon. The P site holds the tRNA attached to the growing polypeptide. The A site holds the tRNA carrying the next amino acid to be added to the polypeptide chain. Discharged tRNA leaves via the E site. Figure 17.16c Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 48. Building a Polypeptide • We can divide translation into three stages – Initiation – Elongation – Termination Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 49. Ribosome Association and Initiation of Translation • The initiation stage of translation – Brings together mRNA, tRNA bearing the first amino acid of the polypeptide, and two subunits of a ribosome t Me P site 3′ U A C 5′ 5′ A U G 3′ Initiator tRNA t Me GTP GDP E mRNA 5′ Start codon mRNA binding site Figure 17.17 3′ Small ribosomal subunit 1 A small ribosomal subunit binds to a molecule of mRNA. In a prokaryotic cell, the mRNA binding site on this subunit recognizes a specific nucleotide sequence on the mRNA just upstream of the start codon. An initiator tRNA, with the anticodon UAC, base-pairs with the start codon, AUG. This tRNA carries the amino acid methionine (Met). Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Large ribosomal subunit 5′ A 3′ Translation initiation complex 2 The arrival of a large ribosomal subunit completes the initiation complex. Proteins called initiation factors (not shown) are required to bring all the translation components together. GTP provides the energy for the assembly. The initiator tRNA is in the P site; the A site is available to the tRNA bearing the next amino acid.
  • 50. Elongation of the Polypeptide Chain • In the elongation stage of translation – Amino acids are added one by one to the preceding amino acid TRANSCRIPTION Amino end of polypeptide DNA mRNA Ribosome TRANSLATION Polypeptide mRNA Ribosome ready for next aminoacyl tRNA E 3′ P A site site 5′ 1 Codon recognition. The anticodon of an incoming aminoacyl tRNA base-pairs with the complementary mRNA codon in the A site. Hydrolysis of GTP increases the accuracy and efficiency of this step. 2 GTP 2 GDP E E P Figure 17.18 3 Translocation. The ribosome translocates the tRNA in the A site to the P site. The empty tRNA in the P site is moved to the E site, where it is released. The mRNA moves along with its bound tRNAs, bringing the next codon to be translated into the A site. P A GDP GTP Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings E P A A 2 Peptide bond formation. An rRNA molecule of the large subunit catalyzes the formation of a peptide bond between the new amino acid in the A site and the carboxyl end of the growing polypeptide in the P site. This step attaches the polypeptide to the tRNA in the A site.
  • 51. Termination of Translation • The final stage of translation is termination – When the ribosome reaches a stop codon in the mRNA Release factor Free polypeptide 5′ 3′ 3′ 5′ 5′ 3′ Stop codon (UAG, UAA, or UGA) 1 When a ribosome reaches a stop 2 The release factor hydrolyzes 3 The two ribosomal subunits codon on mRNA, the A site of the the bond between the tRNA in and the other components of ribosome accepts a protein called the P site and the last amino the assembly dissociate. a release factor instead of tRNA. acid of the polypeptide chain. The polypeptide is thus freed from the ribosome. Figure 17.19 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 52. Polyribosomes • A number of ribosomes can translate a single mRNA molecule simultaneously – Forming a polyribosome Completed polypeptide Growing polypeptides Incoming ribosomal subunits Start of mRNA (5′ end) Polyribosom e End of mRNA (3′ end) (a) An mRNA molecule is generally translated simultaneously by several ribosomes in clusters called polyribosomes. Ribosomes mRNA 0.1 µm Figure 17.20a, b (b) This micrograph shows a large polyribosome in a prokaryotic cell (TEM). Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 53. Completing and Targeting the Functional Protein • Polypeptide chains – Undergo modifications after the translation process Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 54. Protein Folding and Post-Translational Modifications • After translation – Proteins may be modified in ways that affect their three-dimensional shape Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 55. Targeting Polypeptides to Specific Locations • Two populations of ribosomes are evident in cells – Free and bound • Free ribosomes in the cytosol – Initiate the synthesis of all proteins Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 56. • Proteins destined for the endomembrane system or for secretion – Must be transported into the ER – Have signal peptides to which a signalrecognition particle (SRP) binds, enabling the translation ribosome to bind to the ER Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 57. • The signal mechanism for targeting proteins to the ER 1 Polypeptide synthesis begins on a free ribosome in the cytosol. 2 An SRP binds to the signal peptide, halting synthesis momentarily. 3 The SRP binds to a receptor protein in the ER membrane. This receptor is part of a protein complex (a translocation complex) that has a membrane pore and a signal-cleaving enzyme. 4 The SRP leaves, and the polypeptide resumes growing, meanwhile translocating across the membrane. (The signal peptide stays attached to the membrane.) 5 The signalcleaving enzyme cuts off the signal peptide. 6 The rest of the completed polypeptide leaves the ribosome and folds into its final conformation. Ribosome mRNA Signal peptide Signalrecognition particle (SRP) SRP receptor CYTOSOL protein ERLUMEN Translocation complex Figure 17.21 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Signal peptide removed ER membrane Protein
  • 58. • Concept 17.5: RNA plays multiple roles in the cell: a review • RNA – Can hydrogen-bond to other nucleic acid molecules – Can assume a specific three-dimensional shape – Has functional groups that allow it to act as a catalyst Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 59. • Types of RNA in a Eukaryotic Cell Table 17.1 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 60. • Concept 17.6: Comparing gene expression in prokaryotes and eukaryotes reveals key differences • Prokaryotic cells lack a nuclear envelope – Allowing translation to begin while transcription is still in progress RNA polymerase DNA mRNA Polyribosome RNA polymerase Direction of transcription DNA Polyribosome Polypeptide (amino end) Ribosome Figure 17.22 0.25 µm mRNA (5′ end) Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 61. • In a eukaryotic cell – The nuclear envelope separates transcription from translation – Extensive RNA processing occurs in the nucleus Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 62. • Concept 17.7: Point mutations can affect protein structure and function • Mutations – Are changes in the genetic material of a cell • Point mutations – Are changes in just one base pair of a gene Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 63. • The change of a single nucleotide in the DNA’s template strand – Leads to the production of an abnormal protein Wild-type hemoglobin DNA 3′ Mutant hemoglobin DNA C T T G U A 5′ In the DNA, the mutant template strand has an A where the wild-type template has a T. The mutant mRNA has a U instead of an A in one codon. 3′ 5′ T C A mRNA mRNA G A A 5′ 3′ 5′ 3′ Normal hemoglobin Sickle-cell hemoglobin Glu Val Figure 17.23 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings The mutant (sickle-cell) hemoglobin has a valine (Val) instead of a glutamic acid (Glu).
  • 64. Types of Point Mutations • Point mutations within a gene can be divided into two general categories – Base-pair substitutions – Base-pair insertions or deletions Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 65. Substitutions • A base-pair substitution – Is the replacement of one nucleotide and its partner with another pair of nucleotides – Can cause missense or nonsense Wild type mRNA Protein 5′ A U G Met A A G U U U GG C U A A Lys Phe Gly 3′ Stop Amino end Carboxyl end Base-pair substitution No effect on amino acid sequence U instead of C A U G A A G U U U G G U U A A Met Lys Missense Phe Gly Stop A instead of G A U G A A G U U U A G U U A A Met Lys Phe Ser Stop Nonsense U instead of A A U G U A G U U U G G C U A A Figure 17.24 Met Stop Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 66. Insertions and Deletions • Insertions and deletions – Are additions or losses of nucleotide pairs in a gene – May produce frameshift mutations Wild type mRNA Protein 5′ A UG A A GU U U G G C U A A Met Lys Gly Phe Stop Amino end Carboxyl end Base-pair insertion or deletion Frameshift causing immediate nonsense Extra U AU G U A AG U U U G GC U A Met Stop Frameshift causing extensive missense U Missing A U G A A GU U G G C U A A Met Lys Leu Ala Insertion or deletion of 3 nucleotides: no frameshift but extra or missing amino acid A AG Missing A U G U U U G G C U A A Figure 17.25 Met Phe Gly Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Stop 3′
  • 67. Mutagens • Spontaneous mutations – Can occur during DNA replication, recombination, or repair Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 68. • Mutagens – Are physical or chemical agents that can cause mutations Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 69. What is a gene? revisiting the question • A gene – Is a region of DNA whose final product is either a polypeptide or an RNA molecule Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings
  • 70. • A summary of transcription and translation in a eukaryotic cell DNA TRANSCRIPTION 1 RNA is transcribed from a DNA template. 3′ 5′ RNA transcript A lyPo RNA polymerase RNA PROCESSING Exon 2 In eukaryotes, the RNA transcript (premRNA) is spliced and modified to produce mRNA, which moves from the nucleus to the cytoplasm. RNA transcript (pre-mRNA) Intron Aminoacyl-tRNA synthetase p Ca NUCLEUS Amino acid tRNA FORMATION OF INITIATION COMPLEX CYTOPLASM 3 After leaving the nucleus, mRNA attaches to the ribosome. AMINO ACID ACTIVATION 4 Each amino acid attaches to its proper tRNA with the help of a specific enzyme and ATP. Growing polypeptide mRNA A lyPo Activated amino acid Ribosomal subunits p Ca 5′ TRANSLATION C AC U A AC E AAA UG GUU U A U G Codon Figure 17.26 Copyright © 2005 Pearson Education, Inc. publishing as Benjamin Cummings Ribosome A succession of tRNAs add their amino acids to the polypeptide chain Anticodon as the mRNA is moved through the ribosome one codon at a time. (When completed, the polypeptide is released from the ribosome.) 5 A lyPo