The discovery of the genetic role of DNA began with research by Frederick Griffith

in 1928. Griffith worked with two strains of a bacterium, one pathogenic and one
harmless. When he mixed heat-killed remains of the pathogenic strain with living
cells of the harmless strain, some living cells became pathogenic. He called this
phenomenon transformation, now defined as a change in genotype and phenotype due to
assimilation of foreign DNA.

Later work by Oswald Avery, Maclyn McCarty, and Colin MacLeod identified the
transforming substance as DNA, though Many biologists remained skeptical, mainly
because little was known about DNA.

More evidence for DNA as the genetic material came from studies of viruses that
infect bacteria such viruses are called bacteriophages (or phages). A virus is DNA
(sometimes RNA) enclosed by a protective coat, often simply protein – Phages have
been widely used as tools by researchers in molecular genetics.

In 1952, Alfred Hershey and Martha Chase showed that – DNA is the genetic material
of a phage known as T2. They concluded that the injected DNA of the phage provides
the genetic information.

DNA is a polymer of nucleotides, each consisting of a nitrogenous base, a sugar,

and a phosphate group. The nitrogenous bases can be adenine (A), thymine (T),
guanine (G), or cytosine (C).

In 1950, Erwin Chargaff reported that DNA composition varies from one species to
the next, This evidence of molecular diversity among organisms made DNA a more
credible candidate for the genetic material. The basis for these rules was not
understood until the discovery of the double helix.
As such, two findings became known as Chargaff’s rules:
– The base composition of DNA varies between species
– In any species the number of A and T bases is equal and the number of G and C
bases is equal

After DNA was accepted as the genetic material, the challenge was to determine how
its structure accounts for its role in inheritance. Maurice Wilkins and Rosalind
Franklin used a technique called X-ray crystallography to study molecular
structure. Franklin produced a picture of the DNA molecule using this technique.
Franklin’s X-ray crystallographic images of DNA allowed
James Watson to deduce that DNA was helical. The X-ray images also enabled Watson
to deduce the
width of the helix and the spacing of the nitrogenous bases. The pattern in the
photo suggested that the DNA molecule was made up of two strands, forming a double
helix. Franklin had concluded that there were two outer sugar- phosphate backbones,
with the nitrogenous bases paired in the molecule’s interior.

Watson and Crick built models of a double helix to conform to the X-rays and
chemistry of DNA, and then Watson built a model in which the backbones were
antiparallel (their subunits run in opposite directions). At first, Watson and
Crick thought the bases paired like with like (A with A, and so on), but such
pairings did not result in a uniform width. Instead, pairing a purine (A or G) with
a pyrimidine (C or T) resulted in a uniform width consistent with the X-ray data.
The Watson-Crick model explains Chargaff’s rules: in any organism the amount of A =
T, and the amount of G = C.

Resemblance of offspring to parents relies on accurate replication of DNA prior to

meiosis and its transmission to the next generation. Replication prior to mitosis
ensures the faithful transmission of genetic information from a parent cell to the
two daughter cells.
The copying of DNA is called DNA replication. Since the two strands of DNA are
complementary, each strand acts as a template for building a new strand in
replication. This yields two exact replicas of the “parental” molecule.

The copying of DNA is remarkable in its speed and accuracy. More than a dozen
enzymes and other proteins participate in DNA replication. Replication begins at
particular sites called origins of replication, where the two DNA strands are
separated, opening up a replication “bubble”. Replication proceeds in both
directions from each origin, until the entire molecule is copied. An eukaryotic
chromosome may have hundreds or even thousands of origins of replication.

At the end of each replication bubble is a replication fork, a Y-shaped region

where parental DNA strands are being unwound. Helicases are enzymes that untwist
the double helix at the replication forks. Single-strand binding proteins bind to
and stabilize single-stranded DNA. Topoisomerase relieves the strain of twisting of
the double helix by breaking, swiveling, and rejoining DNA strands.

DNA polymerases require a primer to which they can add nucleotides. The initial
nucleotide chain is a short RNA primer. This is synthesized by the enzyme primase.
The completed primer is five to ten nucleotides long. The new DNA strand will start
from the 3′ end of the RNA primer. The antiparallel structure of the double helix
affects replication. DNA polymerases add nucleotides only to the free 3′ end of a
growing strand; therefore, a new DNA strand can elongate only in the 5′ → 3′
direction.

Along one template strand of DNA, the DNA polymerase synthesizes a leading strand
continuously, moving toward the replication fork. o elongate the other new strand,
called the lagging strand, DNA polymerase must work in the direction away from the
replication fork. The lagging strand is synthesized as a series of segments called
Okazaki fragments, which are joined together by DNA ligase.

The proteins that participate in DNA replication form a large complex, a “DNA
replication machine”. The DNA replication machine may be stationary during the
replication process. Recent studies support a model in which DNA polymerase
molecules “reel in” parental DNA and extrude newly made daughter DNA molecules, but
the exact mechanism is not yet resolved.

DNA polymerases proofread newly made DNA, replacing any incorrect nucleotides. DNA
can be damaged by exposure to harmful chemical or physical agents such as cigarette
smoke and X-rays; it can also undergo spontaneous changes. In nucleotide excision
repair, a nuclease cuts out and replaces damaged stretches of DNA. The error rate
after proofreading and repair is low but not zero. Sequence changes may become
permanent and can be passed on to the next generation.

If chromosomes of germ cells became shorter in every cell cycle, essential genes
would eventually be missing from the gametes they produce. An enzyme called
telomerase catalyzes the lengthening of telomeres in germ cells.

The bacterial chromosome is a double-stranded, circular DNA molecule associated

with a small amount of protein. Eukaryotic chromosomes have linear DNA molecules
associated with a large amount of protein. In a bacterium, the DNA is “supercoiled”
and found in a region of the cell called the nucleoid. In the eukaryotic cell, DNA
is precisely combined with proteins in a complex structure called chromatin.
Proteins called histones are responsible for the main level of DNA packing in
interphase chromatin.

Most chromatin is loosely packed in the nucleus during interphase and condenses
prior to mitosis. Loosely packed chromatin is called euchromatin. During interphase
a few regions of chromatin (centromeres and telomeres) are highly condensed into
heterochromatin. Dense packing of the heterochromatin makes it difficult for the
cell to express genetic information coded in these regions.

Histones can undergo chemical modifications that result in changes in chromatin

condensation. These changes can also have multiple effects on gene expression.