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ORGANISATION OF
CHROMOSOMES
INTRODUCTION
   Chromosomes are the structures that contain
    the genetic material
    ◦ They are complexes of DNA and proteins
   The genome comprises all the genetic material
    that an organism possesses
    ◦ In bacteria, it is typically a single circular
      chromosome
    ◦ In eukaryotes, it refers to one complete set of nuclear
      chromosomes
       Note: Eukaryotes possess a mitochondrial genome
              Plants also have a chloroplast genome
Cont…
   The main function of the genetic material is to
    store information required to produce an
    organism
    ◦ The DNA molecule does that through its base
      sequence


   DNA sequences are necessary for
    ◦   1.   Synthesis of RNA and cellular proteins
    ◦   2.   Proper segregation of chromosomes
    ◦   3.   Replication of chromosomes
    ◦   4.   Compaction of chromosomes
         So they can fit within living cells
VIRAL GENOME
   Viruses are small infectious particles containing nucleic acid surrounded
    by a capsid of proteins

   For replication, viruses rely on their host cells
    ◦ ie., the cells they infect

   Most viruses exhibit a limited host range
    ◦ They typically infect only specific types of cells of one host species

   A viral genome is a term used as in whole of the genetic material.
    ◦ Also termed the viral chromosome

   The genome can be
    ◦ DNA or RNA
    ◦ Single-stranded or double-stranded
    ◦ Circular or linear

   Viral genomes vary in size from a few thousand to more than a hundred
    thousand nucleotides
GENERAL STUUCTURE OF VIRUSES
                      Lipid bilayer
                     Picked up when
                  virus leaves host cell
Cont……
 Bacteriophage is very common virus consists of NA+ ptn
  which is usually enclosed by 3 types of capsid structure
 Icosahedral
 Filamentous
 Head and tail

Phage   Host     Shape         Genome Genome No. of
                                      size (kb) genes
MS2     E.coli   Icosahedral   SS linear 3.6    3
                               RNA

M13     E.coli   Filamentous SS linear 6.7      10
                             DNA

T7      E.coli   Head and      DS linear 39.9   55+
                 tail          DNA
Cont……
     Eukaryotic virus consist of capsid + NA
     Capsid can be of Icosahedral and filamentous.
  Virus         Host          Types of    Size of   No. of
                              genome 1    genome    gene
  Parvovirus    Mammals       SS linear   1.6 kb    5
                              DNA
  Tobacco      Plant          SS linear   3.2 kb    4
  mosaic virus                RNA


1- From is given which exist in phage
protein.


Example – HIV virus
BACTERIAL GENOME

    Bacterial chromosomal DNA is usually a
     circular molecule that is a few million
     nucleotides in length.
     ◦ Escherichia coli  ~ 4.6 million base pairs
     ◦ Haemophilus influenzae  ~ 1.8 million base pairs

    A typical bacterial chromosome contains a few
     thousand different genes
     ◦ Structural gene sequences (encoding proteins)
       account for the majority of bacterial DNA
     ◦ The no transcribed DNA between adjacent genes are
       termed intergenic regions
A few hundred
nucleotides in length




                        These play roles in DNA folding, DNA
                        replication, and gene expression
To fit within the bacterial cell, the chromosomal DNA
must be compacted about a 1000-fold This involves
the formation of loop domains. The number of loops
varies according to the size of the bacterial
chromosome and the species.
E. coli has 50-100 with 40,000 to 80,000 bp of DNA in
each.

                             The looped structure compacts
                             the chromosome about 10-fold
   DNA super coiling is a second important way
    to compact the bacterial chromosome

                    Supercoiling within loops creates
                         a more compact DNA




    A SCHEMATIC ILLUSTRATION OF DNA
      SUPER COILING
EUKARYOTIC
CHROMOSOMES
   Eukaryotic species contain one or more sets of
    chromosomes
    ◦ Each set is composed of several different
      linear chromosomes

   The total amount of DNA in eukaryotic species
    is typically greater than that in bacterial cells

   Chromosomes in eukaryotes are located in the
    nucleus
    ◦ To fit in there, they must be highly compacted
       This is accomplished by the binding of
        many proteins
   Eukaryotic genomes vary substantially in size

   In many cases, this variation is not related to
    complexity of the species
    ◦ For example, there is a two fold difference
      in the size of the genome in two closely
      related salamander species.

    ◦ The difference in the size of the genome is
      not because of extra genes
       Rather, the accumulation of repetitive
        DNA sequences
         These do not encode proteins
Has a genome that is more
                  than twice as large as that of




SIZE V/S COMPLEXITY OF THE SPECIES
Organization of Eukaryotic
Chromosomes
   A eukaryotic chromosome contains a long, linear
    DNA molecule

   Three types of DNA sequences are required for
    chromosomal replication and segregation
    ◦ Origins of replication
    ◦ Centromeres
    ◦ Telomeres
A TYPICAL
CHROMATID
DNA to chromosomes
????????????
Eukaryotic Chromatin Compaction
   If stretched end to end, a single set of human
    chromosomes will be over 1 meter long!
       Yet the cell’s nucleus is only 2 to 4 mm in diameter
            Therefore, the DNA must be tightly compacted to fit


   The compaction of linear DNA in eukaryotic
    chromosomes involves interactions between DNA
    and various proteins
       Proteins bound to DNA are subject to change during
        the life of the cell
            These changes affect the degree of chromatin compaction
NUCLEOSOMES
   The repeating structural unit within eukaryotic
    chromatin is the nucleosome

   It is composed of double-stranded DNA
    wrapped around an octamer of histone
    proteins
       An octamer is composed two copies each of four
        different histones
       146 bp of DNA make 1.65 negative superhelical
        turns around the octamer
Vary in length between 20 to 100 bp,
                  depending on species and cell type
                                                             Diameter of the
                                                              nucleosome




   Overall structure of connected nucleosomes resembles “beads on a
    string”
       This structure shortens the DNA length about seven-fold!!!!!!!!!!
   Histone proteins are basic
      They contain many positively-charged amino acids

         Lysine and arginine

      These bind with the phosphates along the DNA

       backbone

   There are five types of histones
      H2A, H2B, H3 and H4 are the core histones

         Two of each make up the octamer




       H1 is the linker histone
          Binds to linker DNA

          Also binds to nucleosomes

              But not as tightly as are the core histones
Play a role in the
 organization and
compaction of the
   chromosome
Nucleosomes Join to Form a 30 nm
Fiber
    Nucleosomes associate with each other to form a more
     compact zig-zag structure fiber of 30 nm. This was
     reveled by F.Thoma in 1977.

    Histone H1 plays a role in this compaction
       At moderate salt concentrations, H1 is removed

          The result is the classic beads-on-a-string

           morphology
       At low salt concentrations, H1 remains bound

          Beads associate together into a more compact

           morphology
   The 30 nm fiber shortens the total length of
    DNA another seven-fold!!!!!!!!!!!!!!!!!!!!!!!!!!!

   Its structure of 30 nm fiber has proven difficult
    to determine
      The DNA conformation may be

        substantially altered when extracted from
        living cells

       Two models have been proposed
          Solenoid model

          Three-dimensional zigzag model
Regular, spiral
    configuration
   containing six
nucleosomes per turn




                               Irregular
                       configuration where
                        nucleosomes have
                         little face-to-face
                                contact
Further Compaction of the
Chromosome
    So far the DNA have been shortened the about 50-
     fold

    A third level of compaction involves interaction
     between the 30 nm fiber and the nuclear matrix

    The nuclear matrix is composed of two parts
        Nuclear lamina
        Internal matrix proteins
             10 nm fiber and associated proteins
SCHEMATIC FIGURE SHOWS THE ARRANGEMENT
OF THE MATRIX WITHIN THE CELL
   The third mechanism of DNA compaction involves
    the formation of radial loop domains



    Matrix-attachment
         regions
          or
                         25,000 to
Scaffold-attachment     200,000 bp
  regions (SARs)




                                        MARs are anchored
                                       to the nuclear matrix,
                                        thus creating radial
                                               loops
Further Compaction of the
Chromosome
     The attachment of radial loops to the nuclear
      matrix is important in two ways
         1. It plays a role in gene regulation

         2. It serves to organize the chromosomes within the
          nucleus
              Each chromosome in the nucleus is located in a discrete and
               nonoverlapping chromosome territory
Heterochromatin vs
         Euchromatin
   The compaction level of interphase chromosomes is not
    completely uniform (German cytologist E. Heitz in 1928)
      Euchromatin

            Less condensed regions of chromosomes
            Transcriptionally active
            Regions where 30 nm fiber forms radial loop domains


       Heterochromatin
            Tightly compacted regions of chromosomes
            Transcriptionally inactive (in general)
            Radial loop domains compacted even further
   There are two types of heterochromatin
       Constitutive heterochromatin
            Regions that are always heterochromatic
            Permanently inactive with regard to transcription
       Facultative heterochromatin
            Regions that can interconvert between euchromatin and
             heterochromatin
            Example: Barr body
Compaction level
                     in euchromatin



During interphase
most chromosomal     Compaction level
   regions are      in heterochromatin
  euchromatic
The number of loops has not changed
                      However, the diameter of each loop is smaller

During interphase,
condensin is in the
cytoplasm




                                                                      Condesin binds to
                                                                      chromosomes and
                                                                        compacts the
                                                                         radial loops




                            Condesin travels
                            into the nucleus




THE CONDENSATION OF A METAPHASE
CHROMOSOME BY CONDENSIN
Metaphase Chromosomes
   As cells enter M phase, the level of compaction changes
    dramatically
      By the end of prophase, sister chromatids are entirely

       heterochromatic
      Two parallel chromatids have an overall diameter of

       1,400 nm

   These highly condensed metaphase chromosomes
    undergo little gene transcription
   In metaphase chromosomes the radial loops are highly
    compacted and stay anchored to a scaffold
      The scaffold is formed from the nuclear matrix




   Histones are needed for the compaction of radial loops
   Two multiprotein complexes help to form and organize
    metaphase chromosomes

       Condensin
          Plays a critical role in chromosome condensation




       Cohesin
          Plays a critical role in sister chromatid alignment




   Both contain a category of proteins called SMC proteins
      Acronym = Structural maintenance of chromosomes

      SMC proteins use energy from ATP and catalyze

       changes in chromosome structure
       lets view the process of chromosomes
        organizations once again..!!!!!!!!!!!!!!!!!!!
Excuse me, your mess during DNA
remolding….!!!!!!!!!!!!



THANK U FOR UR CO- OPERATION

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organization of DNA in chromosomes.

  • 2. INTRODUCTION  Chromosomes are the structures that contain the genetic material ◦ They are complexes of DNA and proteins  The genome comprises all the genetic material that an organism possesses ◦ In bacteria, it is typically a single circular chromosome ◦ In eukaryotes, it refers to one complete set of nuclear chromosomes  Note: Eukaryotes possess a mitochondrial genome Plants also have a chloroplast genome
  • 3. Cont…  The main function of the genetic material is to store information required to produce an organism ◦ The DNA molecule does that through its base sequence  DNA sequences are necessary for ◦ 1. Synthesis of RNA and cellular proteins ◦ 2. Proper segregation of chromosomes ◦ 3. Replication of chromosomes ◦ 4. Compaction of chromosomes  So they can fit within living cells
  • 4. VIRAL GENOME  Viruses are small infectious particles containing nucleic acid surrounded by a capsid of proteins  For replication, viruses rely on their host cells ◦ ie., the cells they infect  Most viruses exhibit a limited host range ◦ They typically infect only specific types of cells of one host species  A viral genome is a term used as in whole of the genetic material. ◦ Also termed the viral chromosome  The genome can be ◦ DNA or RNA ◦ Single-stranded or double-stranded ◦ Circular or linear  Viral genomes vary in size from a few thousand to more than a hundred thousand nucleotides
  • 5. GENERAL STUUCTURE OF VIRUSES Lipid bilayer Picked up when virus leaves host cell
  • 6. Cont……  Bacteriophage is very common virus consists of NA+ ptn which is usually enclosed by 3 types of capsid structure  Icosahedral  Filamentous  Head and tail Phage Host Shape Genome Genome No. of size (kb) genes MS2 E.coli Icosahedral SS linear 3.6 3 RNA M13 E.coli Filamentous SS linear 6.7 10 DNA T7 E.coli Head and DS linear 39.9 55+ tail DNA
  • 7. Cont……  Eukaryotic virus consist of capsid + NA  Capsid can be of Icosahedral and filamentous. Virus Host Types of Size of No. of genome 1 genome gene Parvovirus Mammals SS linear 1.6 kb 5 DNA Tobacco Plant SS linear 3.2 kb 4 mosaic virus RNA 1- From is given which exist in phage protein. Example – HIV virus
  • 8. BACTERIAL GENOME  Bacterial chromosomal DNA is usually a circular molecule that is a few million nucleotides in length. ◦ Escherichia coli  ~ 4.6 million base pairs ◦ Haemophilus influenzae  ~ 1.8 million base pairs  A typical bacterial chromosome contains a few thousand different genes ◦ Structural gene sequences (encoding proteins) account for the majority of bacterial DNA ◦ The no transcribed DNA between adjacent genes are termed intergenic regions
  • 9. A few hundred nucleotides in length These play roles in DNA folding, DNA replication, and gene expression
  • 10. To fit within the bacterial cell, the chromosomal DNA must be compacted about a 1000-fold This involves the formation of loop domains. The number of loops varies according to the size of the bacterial chromosome and the species. E. coli has 50-100 with 40,000 to 80,000 bp of DNA in each. The looped structure compacts the chromosome about 10-fold
  • 11. DNA super coiling is a second important way to compact the bacterial chromosome Supercoiling within loops creates a more compact DNA A SCHEMATIC ILLUSTRATION OF DNA SUPER COILING
  • 12. EUKARYOTIC CHROMOSOMES  Eukaryotic species contain one or more sets of chromosomes ◦ Each set is composed of several different linear chromosomes  The total amount of DNA in eukaryotic species is typically greater than that in bacterial cells  Chromosomes in eukaryotes are located in the nucleus ◦ To fit in there, they must be highly compacted  This is accomplished by the binding of many proteins
  • 13. Eukaryotic genomes vary substantially in size  In many cases, this variation is not related to complexity of the species ◦ For example, there is a two fold difference in the size of the genome in two closely related salamander species. ◦ The difference in the size of the genome is not because of extra genes  Rather, the accumulation of repetitive DNA sequences  These do not encode proteins
  • 14. Has a genome that is more than twice as large as that of SIZE V/S COMPLEXITY OF THE SPECIES
  • 15. Organization of Eukaryotic Chromosomes  A eukaryotic chromosome contains a long, linear DNA molecule  Three types of DNA sequences are required for chromosomal replication and segregation ◦ Origins of replication ◦ Centromeres ◦ Telomeres
  • 18. Eukaryotic Chromatin Compaction  If stretched end to end, a single set of human chromosomes will be over 1 meter long!  Yet the cell’s nucleus is only 2 to 4 mm in diameter  Therefore, the DNA must be tightly compacted to fit  The compaction of linear DNA in eukaryotic chromosomes involves interactions between DNA and various proteins  Proteins bound to DNA are subject to change during the life of the cell  These changes affect the degree of chromatin compaction
  • 19. NUCLEOSOMES  The repeating structural unit within eukaryotic chromatin is the nucleosome  It is composed of double-stranded DNA wrapped around an octamer of histone proteins  An octamer is composed two copies each of four different histones  146 bp of DNA make 1.65 negative superhelical turns around the octamer
  • 20. Vary in length between 20 to 100 bp, depending on species and cell type Diameter of the nucleosome  Overall structure of connected nucleosomes resembles “beads on a string”  This structure shortens the DNA length about seven-fold!!!!!!!!!!
  • 21. Histone proteins are basic  They contain many positively-charged amino acids  Lysine and arginine  These bind with the phosphates along the DNA backbone  There are five types of histones  H2A, H2B, H3 and H4 are the core histones  Two of each make up the octamer  H1 is the linker histone  Binds to linker DNA  Also binds to nucleosomes  But not as tightly as are the core histones
  • 22. Play a role in the organization and compaction of the chromosome
  • 23. Nucleosomes Join to Form a 30 nm Fiber  Nucleosomes associate with each other to form a more compact zig-zag structure fiber of 30 nm. This was reveled by F.Thoma in 1977.  Histone H1 plays a role in this compaction  At moderate salt concentrations, H1 is removed  The result is the classic beads-on-a-string morphology  At low salt concentrations, H1 remains bound  Beads associate together into a more compact morphology
  • 24. The 30 nm fiber shortens the total length of DNA another seven-fold!!!!!!!!!!!!!!!!!!!!!!!!!!!  Its structure of 30 nm fiber has proven difficult to determine  The DNA conformation may be substantially altered when extracted from living cells  Two models have been proposed  Solenoid model  Three-dimensional zigzag model
  • 25. Regular, spiral configuration containing six nucleosomes per turn Irregular configuration where nucleosomes have little face-to-face contact
  • 26. Further Compaction of the Chromosome  So far the DNA have been shortened the about 50- fold  A third level of compaction involves interaction between the 30 nm fiber and the nuclear matrix  The nuclear matrix is composed of two parts  Nuclear lamina  Internal matrix proteins  10 nm fiber and associated proteins
  • 27. SCHEMATIC FIGURE SHOWS THE ARRANGEMENT OF THE MATRIX WITHIN THE CELL
  • 28. The third mechanism of DNA compaction involves the formation of radial loop domains Matrix-attachment regions or 25,000 to Scaffold-attachment 200,000 bp regions (SARs) MARs are anchored to the nuclear matrix, thus creating radial loops
  • 29. Further Compaction of the Chromosome  The attachment of radial loops to the nuclear matrix is important in two ways  1. It plays a role in gene regulation  2. It serves to organize the chromosomes within the nucleus  Each chromosome in the nucleus is located in a discrete and nonoverlapping chromosome territory
  • 30. Heterochromatin vs Euchromatin  The compaction level of interphase chromosomes is not completely uniform (German cytologist E. Heitz in 1928)  Euchromatin  Less condensed regions of chromosomes  Transcriptionally active  Regions where 30 nm fiber forms radial loop domains  Heterochromatin  Tightly compacted regions of chromosomes  Transcriptionally inactive (in general)  Radial loop domains compacted even further
  • 31. There are two types of heterochromatin  Constitutive heterochromatin  Regions that are always heterochromatic  Permanently inactive with regard to transcription  Facultative heterochromatin  Regions that can interconvert between euchromatin and heterochromatin  Example: Barr body
  • 32.
  • 33. Compaction level in euchromatin During interphase most chromosomal Compaction level regions are in heterochromatin euchromatic
  • 34. The number of loops has not changed However, the diameter of each loop is smaller During interphase, condensin is in the cytoplasm Condesin binds to chromosomes and compacts the radial loops Condesin travels into the nucleus THE CONDENSATION OF A METAPHASE CHROMOSOME BY CONDENSIN
  • 35. Metaphase Chromosomes  As cells enter M phase, the level of compaction changes dramatically  By the end of prophase, sister chromatids are entirely heterochromatic  Two parallel chromatids have an overall diameter of 1,400 nm  These highly condensed metaphase chromosomes undergo little gene transcription  In metaphase chromosomes the radial loops are highly compacted and stay anchored to a scaffold  The scaffold is formed from the nuclear matrix  Histones are needed for the compaction of radial loops
  • 36. Two multiprotein complexes help to form and organize metaphase chromosomes  Condensin  Plays a critical role in chromosome condensation  Cohesin  Plays a critical role in sister chromatid alignment  Both contain a category of proteins called SMC proteins  Acronym = Structural maintenance of chromosomes  SMC proteins use energy from ATP and catalyze changes in chromosome structure  lets view the process of chromosomes organizations once again..!!!!!!!!!!!!!!!!!!!
  • 37.
  • 38. Excuse me, your mess during DNA remolding….!!!!!!!!!!!! THANK U FOR UR CO- OPERATION