DNA Replication

Definition:
“DNA replication is a biological process of producing two identical replicas of DNA from one
original DNA molecule.”

Structure of DNA
DNA exists as a double-stranded structure, with both strands coiled together to form the
characteristic double helix. DNA strands have a directionality, and the different ends of a single
strand are called “3’ (three-prime) end” and “5’ (five-prime) end”.
DNA is made up of a double helix of two complementary strands. During replication, these
strands are separated. Each strand of the original DNA molecule then serves as a template for the
production of its counterpart, a process referred to as semiconservative replication.

Modes of Replication
Three modes of DNA replication have been reported which are
i. Semiconservative Replication
ii. Conservative Replication
iii. Dispersive Replication

1. Semiconservative Replication
In the semiconservative model, the two parental strands separate and each makes a copy of itself.
After one round of replication, the two daughter molecules each comprises one old and one new
strand. Note that after two rounds, two of the DNA molecules consist only of new material,
while the other two contain one old and one new strand.

Semiconservative Model
2. Conservative Replication
In the conservative model, the parental molecule directs synthesis of an entirely new double-
stranded molecule, such that after one round of replication, one molecule is conserved as two old
strands. This is repeated in the second round.

Conservative Model

3. Dispersive Replication
In the dispersive model, material in the two parental strands is distributed more or less randomly
between two daughter molecules. In the model shown here, old material is distributed
symmetrically between the two daughter molecules. Other distribution are possible.

Dispersive Model
DNA replication is having three major steps

1.Initiation

DNA synthesis is initiated at particular points within the DNA strand known as ‘origins’,
which are specific coding regions. These origins are targeted by initiator proteins, which go
on to recruit more proteins that help aid the replication process, forming a replication
complex around the DNA origin. There are multiple origin sites, and when replication of
DNA begins, these sites are referred to as Replication Forks.

● Within the replication complex is the enzyme DNA Helicase, which unwinds the double
helix and exposes each of the two strands, so that they can be used as a template for
replication.
 ● DNA can only be extended via the addition of a free nucleotide triphosphate to the 3’-
end of a chain. As the double helix runs antiparallel, but DNA replication only occurs in
one direction.

● DNA Primase is another enzyme that is important in DNA replication. It synthesises a

small RNA primer, which acts as a ‘kick-starter’ for DNA Polymerase.

Elongation:
During elongation in DNA replication, DNA polymerase starts adding new nucleotides to the 3’
end of newly synthesized polynucleotide strand. The template strand specifies which of the four
DNA nucleotides (adenine, thymine, cytosine or guanine) is added at each position along the
new chain. Only the nucleotide complimentary to the template nucleotide at that positon is added
to the new strand.

DNA polymerase:

DNA polymerase is an enzyme that builds a new duplex DNA strand by adding new nucleotides
in the 5’ to 3’ direction. Also performs proof-reading and detects error in the pattern. It uses one
strand as a template. DNA polymerase requires a primer, which is complementary to the
template.

Types of DNA polymerases:

There are three kinds of polymerases that are:

1. DNA polymerase I
2. DNA polymerase II
3. DNA polymerase III
All of these three enzymes are used in replication and repair of the DNA. DNA polymerases I
proof reads the DNA and replaces the RNA primer with DNA. DNA polymerases III synthesize
new strands of DNA. DNA polymerase contain s a groove that allows it to bind to a single
stranded template DNA and travel one nucleotide at one time.

DNA polymerase can’t initiate new strand synthesis; it only adds new nucleotides at the 3’ end
of existing strands.
Primase

All newly synthesized polynucleotide strands must be initiated by a specialized RNA

polymerase called primase. Primase initiates polynucleotide synthesis and by creating a short
RNA polynucleotide strand complementary to template DNA strand. Eventually the RNA
nucleotides in the primer are removed and replaced with the DNA nucleotides.

The Leading Strands and Lagging Strands:

These two newly synthesized strands grown in opposite directions because the template strands
at each replication fork are antiparallel. The leading strand is synthesized continuously towards
the replication fork as helicase unwinds the template double-stranded DNA.

The lagging strand is synthesized in the direction away from the replication fork and away from
the DNA helicase unwinds. This lagging strand is synthesized in pieces because the DNA
polymerase can only synthesize in the 5’ to 3’ direction, and so it constantly encounters the
previously synthesized new strand

Okazaki Fragments:

The leading strand is one complete strand, while the lagging strand is not. It is instead made up
of multiple ‘mini strands’, known as “Okazaki fragments”.
TERMINATION:
The process of expanding the new DNA strands continues until there is either no more DNA
template left to replicate or two replication forks meet and subsequently terminate.

Enzymes:

Enzymes that are involve in the termination are:

● Exonuclease
● DNA ligase
● Telomerase