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S.Kaayathri Devi
I6UTIO
III B.Sc., Biotechnology
ANJAC
Central dogma of biology
REPLICATION
DNA DNA
TRANSCRIPTION POST
TRANSCRIPTIONAL
MODIFICATION
RNA
TRANSLATION POST TRANSLATIONAL
MODIFICATION
PROTEIN
INTRODUCTION
 Synthesis of RNA by copying DNA template is known
as transcription.
 Only selected portion of the genome (exons)is
transcribed .
 RNA polymerase requires no primers it is a DNA
dependent RNA polymerase
The process of transcription takes place in three major
steps. They are :
1. Initiation.
2.Elongation.
3.Termination.
INITIATIATION
 Transcription is initiated by RNA Polymerase
holoenzyme from a specific region called promotor
sequence in DNA .
 Principle enzyme – Bacterial RNA Polymerase
 RNA Polymerase alone is called core Polymerase .
 Core polymerase consists of α,β,β’and ω sub units.
The binding of core polymerase to promotor
sequence is facilated by σ factor.
Core polymerase + σ factor = RNA polymerase
holoenzyme
Promotor sequence E. Coli :
1. 5’-TTGACA-3’ at – 35 element
(pribnow box)
2. 5’ – TATAAT – 3’ at – 10 element
(TATAA box)
 Closed complex formation :
Binding of RNA polymerase holoenzyme
to promotor sequence.
 Open complex formation :
1. RNA polymerase holoenzyme separates
10-14 DNA basepairs extending from
-11 to +3 and this is called melting.
2. It leads to the formation of open
complex.
3. closed complex to open complex
isomerization
Teritiary complex formation :
1. If the enzyme synthesizes RNA <10
basepairs ,it does not elongate it
abortive initiation.
2. Because σ factor acts as a mimic of RNA
and blocks the RNA exit channel in open complex.
3. When RNA polymerase synthesizes
RNA > 10 base pairs , it ejects σ factor and RNA
elongates further and moves out through exit channel.
This is known as the teritiary complex
ELONGATION
 After the formation of teritiary complex the RNA
elongates further by the addition of dNTPs .
 The synthesized is proof read by hydrolytic editing.
Gre factor enhances the proof reading activity.
Pyrophospholytic editing is another proof reading
mechanism.
TERMINATION
 The process of termination can happen in two
different mechanisms. They are :
1. Rho – independent termination.
2. Rho – dependent termination.
Rho – independent termination
In this mechanism, transcription is terminated by
termination sequence in DNA.
Terminator sequence contains invert repeat causing
complementary pairing as transcript RNA form
hairpin structure.
Invert repeat is followed by a large number of T (~8
basepairs), in RNA “U” appears.
The load of hairpin structure isn’t tolerated by A=U
bps so RNA gets separated from RNA –DNA
heteroduplex.
Rho – dependent termination
In this mechanism,transcription is terminated by rho
protein (ρ).
Rho protein is a single strand ATPase protein.
Rho protein binds to the single stranded RNA after
termination.
In bacteria, transcription and translation occurs
simultaneously so Rho protein binds to the mRNA
after translation i.e.,when translation is completed but
transcription is “ON”.

Transcription in prokaryotes

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Transcription in prokaryotes

  • 2. Central dogma of biology REPLICATION DNA DNA TRANSCRIPTION POST TRANSCRIPTIONAL MODIFICATION RNA TRANSLATION POST TRANSLATIONAL MODIFICATION PROTEIN
  • 3.
  • 4. INTRODUCTION  Synthesis of RNA by copying DNA template is known as transcription.  Only selected portion of the genome (exons)is transcribed .  RNA polymerase requires no primers it is a DNA dependent RNA polymerase
  • 5.
  • 6. The process of transcription takes place in three major steps. They are : 1. Initiation. 2.Elongation. 3.Termination.
  • 7. INITIATIATION  Transcription is initiated by RNA Polymerase holoenzyme from a specific region called promotor sequence in DNA .  Principle enzyme – Bacterial RNA Polymerase  RNA Polymerase alone is called core Polymerase .  Core polymerase consists of α,β,β’and ω sub units.
  • 8.
  • 9. The binding of core polymerase to promotor sequence is facilated by σ factor. Core polymerase + σ factor = RNA polymerase holoenzyme Promotor sequence E. Coli : 1. 5’-TTGACA-3’ at – 35 element (pribnow box) 2. 5’ – TATAAT – 3’ at – 10 element (TATAA box)
  • 10.  Closed complex formation : Binding of RNA polymerase holoenzyme to promotor sequence.  Open complex formation : 1. RNA polymerase holoenzyme separates 10-14 DNA basepairs extending from -11 to +3 and this is called melting. 2. It leads to the formation of open complex. 3. closed complex to open complex isomerization
  • 11. Teritiary complex formation : 1. If the enzyme synthesizes RNA <10 basepairs ,it does not elongate it abortive initiation. 2. Because σ factor acts as a mimic of RNA and blocks the RNA exit channel in open complex. 3. When RNA polymerase synthesizes RNA > 10 base pairs , it ejects σ factor and RNA elongates further and moves out through exit channel. This is known as the teritiary complex
  • 12.
  • 13.
  • 14.
  • 15. ELONGATION  After the formation of teritiary complex the RNA elongates further by the addition of dNTPs .  The synthesized is proof read by hydrolytic editing. Gre factor enhances the proof reading activity. Pyrophospholytic editing is another proof reading mechanism.
  • 16.
  • 17. TERMINATION  The process of termination can happen in two different mechanisms. They are : 1. Rho – independent termination. 2. Rho – dependent termination.
  • 18. Rho – independent termination In this mechanism, transcription is terminated by termination sequence in DNA. Terminator sequence contains invert repeat causing complementary pairing as transcript RNA form hairpin structure. Invert repeat is followed by a large number of T (~8 basepairs), in RNA “U” appears. The load of hairpin structure isn’t tolerated by A=U bps so RNA gets separated from RNA –DNA heteroduplex.
  • 19.
  • 20. Rho – dependent termination In this mechanism,transcription is terminated by rho protein (ρ). Rho protein is a single strand ATPase protein. Rho protein binds to the single stranded RNA after termination. In bacteria, transcription and translation occurs simultaneously so Rho protein binds to the mRNA after translation i.e.,when translation is completed but transcription is “ON”.
  • 21.
  • 22.