Week XI

• Chromatin structure and nucleosomes

• Heterochromatin
• Satellite DNA
• Centromeric DNA sequences
• DNA is organized in eukaryotic chromosomes, which are most clearly
visible as highly condensed structures during mitosis
• After chromosome separation and cell division, cells enter the
interphase stage of the cell cycle, at which time the components
of the chromosome uncoil and decondense into a form referred to as
chromatin
• While in interphase, the chromatin is dispersed throughout the
nucleus
• Histones contain large amounts of the positively charged amino acids
lysine and arginine, making it possible for them to bond
electrostatically to the negatively charged phosphate groups of
nucleotides
• The general model for chromatin structure is based on the
assumption that chromatin fibers, composed of DNA and protein,
undergo extensive coiling and folding as they are condensed within
the cell nucleus
• Chromatin produces regularly spaced diffraction rings, suggesting that
repeating structural units occur along the chromatin axis
• If the histone molecules are chemically removed from chromatin,
the regularity of this diffraction pattern is disrupted
• Digestion of chromatin by certain endonucleases, such as micrococcal
nuclease, yields DNA fragments that are approximately 200 base pairs
in length
• Thus, chromatin consists of some type of repeating unit, each of
which protects the DNA from enzymatic cleavage except where any
two units are joined
• It is the area between units that is attacked and cleaved by the
endonuclease
• Electron microscopic observations of
chromatin have revealed that
chromatin fibers are composed of
linear arrays of spherical particles
• Discovered by Ada and Donald Olins,
the particles occur regularly
along the axis of a chromatin strand
and resemble
• These particles, initially referred to as
v-bodies (v is the Greek letter nu), are
now called nucleosomes
• When nuclease digestion time is extended, some of the 200 base
pairs of DNA are removed from the nucleosome, creating what is
called a nucleosome core particle consisting of 147 base pairs
• The DNA lost in the prolonged digestion is responsible for
linking nucleosomes together
• This linker DNA is associated with the fifth histone, H1
Relatively relaxed chromatin – Transcriptionally active
Condensed – Transcriptionally inactive
Heterochromatin

• In the early part of the twentieth century, it was observed that some
parts of the chromosome remain condensed and stain deeply during
interphase, while most parts are partially uncoiled and do not stain
• In 1928, the terms euchromatin and heterochromatin were coined to
describe the parts of chromosomes that are uncoiled and those that
remain condensed, respectively
• Heterochromatic areas are genetically inactive because they either
lack genes or contain genes that are repressed
• Heterochromatin replicates later during the S phase of the cell cycle
than does euchromatin
Heterochromatin

• The discovery of heterochromatin provided the first clues that parts

of eukaryotic chromosomes do not always encode proteins
• For example, one particular heterochromatic region of the
chromosome, the telomere, is thought to be involved in maintenance
of the chromosome’s structural integrity, and another region, the
centromere, is involved in chromosome movement during cell division
Position effect
• When certain heterochromatic areas from one chromosome are
translocated to a new site on the same or another nonhomologous
chromosome, genetically active areas sometimes become genetically
inert if they now lie adjacent to the translocated heterochromatin
• This influence on existing euchromatin is one example of what is
more generally referred to as a position effect
• That is, the position of a gene or group of genes relative to all other
genetic material may affect their expression
Satellite DNA

• They are highly repetitive DNA consisting

of short sequences repeated several
times
• It carries a variable AT-rich repeat unit
that often forms arrays up to 100 mb
• The monomer length of satDNA
sequences ranges from 150-400 bp in the
majority of plants and animals
• Satelite DNA was first characterized from
an Ultra centrifugation reaction
• Main genomic DNA forms thick band and
small
Satellite DNA

• They have less GC content than main

• Can be found in different regions, telomeres and centromeres
• Satellite DNA (satDNA) is the highly repetitive DNA consisting of short
sequence repeated a large number of times
• It carries a variable AT-rich repeat unit that often forms arrays up to
100 Mb
Satellite DNA​

• Minisatellites are short tandem repeats of DNA sequences, typically

consisting of repeats that are 10 to 100 base pairs long
• Minisatellites are often found at telomeres (the ends of chromosomes) and
can also be dispersed throughout the genome
• Microsatellites (or Short Tandem Repeats - STRs) are even shorter tandem
repeats, usually consisting of repeats that are 2 to 6 base pairs long
• They are widely distributed throughout the genome, both in coding and
non-coding regions
• Microsatellites are commonly used in DNA profiling and genetic
fingerprinting due to their variability among individuals
Satellite DNA

• The functions of satellite DNA are not entirely clear, and their
presence in the genome may not be directly related to coding for
proteins
• They have been associated with structural roles in the organization of
chromatin and chromosomes
• Additionally, variations in the length of microsatellites among
individuals contribute to genetic diversity and are useful in genetic
studies, forensics, and paternity testing
Centromeric DNA sequences

• Centromeric DNA sequences are specific DNA sequences found at the

centromere of a eukaryotic chromosome
• The centromere is a specialized region on a chromosome that plays a
crucial role in cell division, particularly in the segregation of
chromosomes during mitosis and meiosis
• Centromeres are essential for proper chromosome movement and
distribution to daughter cells
Centromeric DNA sequences

• The primary function of

centromeres is to provide a site for
the assembly of the kinetochore, a
protein complex that interacts with
spindle fibers during cell division
• The kinetochore facilitates the
attachment of chromosomes to the
mitotic spindle and ensures
accurate segregation of genetic
material
• In humans, common centromeric
repeat is the alpha satellite DNA
Centromeric DNA sequences

• Point Centromeres Found in organisms like yeast, these centromeres

are relatively small and consist of a specific DNA sequence where the
kinetochore assembles
• Regional or Holocentric Centromeres Found in some plants and
invertebrates, these centromeres are larger and cover a broader
region of the chromosome