The proteins that bind to the DNA to form eukaryotic chromosomes are

traditionally divided into two general classes: (1) basic (positively charged
at neutral pH) proteins called histones and (2) a heterogeneous, largely
acidic (negatively charged at neutral pH) group of proteins collectively
referred to as nonhistone chromosomal proteins. The complex of both
classes of protein with the nuclear DNA of eukaryotic cells is known as
chromatin. Histones play a major structural role in chromatin. They are
present in the chromatin of all eukaryotes in such enormous quantities in
the cell (about 60 million molecules of each type per human
cell) that their total mass in chromatin is about equal to that of the DNA.
Histones are responsible for the first and most basic level of chromosome
packing, the nucleosome. The nonhistone chromosomal proteins probably
do not play central roles in the packaging of DNA into chromosomes.
Instead, they are likely candidates for roles in regulating the expression of
specific genes or sets of genes.
When interphase nuclei are broken open very gently and their contents
examined under the electron microscope, most of the chromatin is in the
form of a fibre with a diameter of about 30 nm. If this chromatin is
subjected to treatments that cause it to unfold partially, it can be seen
under the electron microscope as a series of "beads on a string". The
string is DNA, and each bead is a "nucleosome core particle" that consists
of DNA wound around a protein core formed from histones.
The structural organization of nucleosomes was determined after first
isolating them from unfolded chromatin by digestion with particular
enzymes (call endonucleases) that break down DNA by cutting between
the nucleosomes. After digestion for a short period, the exposed DNA
between the nucleosome core particles, the linker DNA, is degraded. Each
individual nucleosome core particle consists of a complex of eight histone
proteins-two molecules each of histones H2A, H2B, H3, and H4-and
double-stranded DNA that is 147 nucleotide pairs long. The histone
octamer forms a protein core around which the double-stranded DNA is
wound. Each nucleosome core particle is separated from the next by a
region of linker DNA, which can vary in length from a few nucleotide pairs
up to about 80. (The term nucleosome technically refers to a nucleosome
core
particle
plus
one of
its

adjacent DNA linkers, but it is often used synonymously with nucleosome

core particle.) On average therefore nucleosomes repeat at intervals of
about 200 nucleotide pairs.

1. Digestion of chromatin by certain endonucleases, such as micrococcal

nuclease, yields DNA fragments that are approximately 200 base pairs in
length.
2. Electron microscopic observations of chromatin have revealed that
chromatin fibres are composed of linear arrays of spherical particles. The
particles occur regularly along the axis of a chromatin strand and
resemble beads on a string. These particles initially referred to as
nucleosomes.
3. Studies of the chemical association between histone molecules and
DNA in the nucleosomes of chromatin show that histones H2A, H2B, H3,
and H4 occur as two types of tetramers, (H2A) 2 (H2B) 2 and (H3) 2
(H4) 2. Each repeating nucleosome unit consists of an octamer in
association with about 200 base pairs of DNA.
4. 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.

In this model, a 147-bp length of the 2-nm-diameter DNA molecule coils

around an octamer of histones in a left-handed super helix that wrap
about 1.7 turns per nucleosome.
Each nucleosome, ellipsoidal in shape, measures about (approximately 11
nm in diameter by 6.5 nm high).
Significantly, the formation of the nucleosome represents the first level of
packing, whereby the DNA helix is reduced to about one-third of its
original length by winding around the histones. The 11-nmdiameter fibre
is further packed into a thicker, 30-nm-diameter structure that was initially
called a solenoid.
This thicker structure, which is dependent on the presence of histone H1,
consists of numerous nucleosomes coiled around and stacked upon one
another, creating a second level of packing. This provides a six-fold
increase in compaction of the DNA. The 30-nm structures are folded into a

series of looped domains, which further condense the chromatin fibre into
a structure that is 300 nm in diameter.
These coiled chromatin fibres are then compacted into the chromosome
arms that constitute a chromatid, one of the longitudinal subunits of the
metaphase chromosome. While the chromatid arms to be 700 nm in
diameter, this value undoubtedly varies among different organisms. At a
value of 700 nm, a pair of sister chromatids comprising a chromosome
measures about 1400 nm.