DNA REPLICATION AND

PROTEIN SYNTHESIS
Why Replicate DNA?
• DNA is the genetic material that defines every cell. Before a cell
duplicates and is divided into new daughter cells through either
mitosis or meiosis, biomolecules and organelles must be copied to
be distributed among the cells. DNA, found within the nucleus, must
be replicated in order to ensure that each new cell receives the
correct number of chromosomes. The process of DNA duplication is
called DNA replication. Replication follows several steps that
involve multiple proteins called replication enzymes and RNA. In
eukaryotic cells, such as animal cells and plant cells, DNA replication
occurs in the S phase of interphase during the cell cycle. The
process of DNA replication is vital for cell growth, repair, and
reproduction in organisms.
DNA Replication
• In molecular biology, DNA replication is the biological
process of producing two identical replicas of DNA from
one original DNA molecule. DNA replication occurs in all
living organisms acting as the most essential part of
biological inheritance.
• This is essential for cell division during growth and repair of
damaged tissues, while it also ensures that each of the new
cells receives its own copy of the DNA. The cell possesses
the distinctive property of division, which makes
replication of DNA essential.
Summary of DNA Replication
• DNA replication can be thought of in three stages: initiation, elongation and termination
Initiation
• DNA synthesis is initiated at particular points within the DNA strand known as ‘origins’, which
have 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. Multiple origin sites exist within the DNA’s structure; when replication of DNA
begins, these sites are referred to as replication forks.
• Within the replication complex is the DNA helicase. This enzyme unwinds the double helix and
exposes each of the two strands so that they can be used as a template for replication. It does
this by hydrolysing the ATP used to form the bonds between the nucleobases, thereby breaking
the bond holding the two strands together.
• 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. This enzyme is ultimately responsible
for the creation and expansion of new strands of DNA.
Elongation
• Once DNA Polymerase has attached to the two unzipped strands of DNA (i.e. the template
strands), it is able to start synthesising new strands of DNA to match the templates. DNA
polymerase is only able to extend the primer by adding free nucleotides to the 3’ end.
• One of the template strands is read in a 3’ to 5’ direction, therefore the new strand will be
formed in a 5’ to 3’ direction. This newly formed strand is referred to as the leading strand.
Along the leading strand, DNA primase only needs to synthesise an RNA primer once, at the
beginning, to initiate DNA polymerase. This is because DNA polymerase is able to extend the
new DNA strand by reading the template 3′ to 5′, synthesising in a 5 ′ to 3 ′ direction as noted
above.
• However, the other template strand (the lagging strand) is antiparallel and is therefore read in
a 5’ to 3’ direction. Continuous DNA synthesis, as in the leading strand, would need to be in
the 3′ to 5′ direction, which is impossible as DNA polymerase cannot add bases to the 5 ′ end.
Instead, as the helix unwinds, RNA primers are added to the newly exposed bases on the
lagging strand and DNA synthesis occurs in fragments, but still in the 5′ to 3′ direction as
before. These fragments are known as Okazaki fragments.
 Termination
• The process of expanding the new DNA strands continues until there is either no more
DNA template strand left to replicate (i.e. at the end of the chromosome) or two
replication forks meet and subsequently terminate. The meeting of two replication
forks is not regulated and happens randomly along the course of the chromosome.
• Once DNA synthesis has finished, the newly synthesised strands are bound and
stabilised. For the lagging strand, two enzymes are needed to achieve this
stabilisation: Helicase removes the RNA primer at the beginning of each Okazaki
fragment, and DNA ligase joins these fragments together to create one complete
strand.
• DNA is made up of a double helix of two
complementary strands. The double helix
describes the appearance of a double-
stranded DNA which is thus composed of
two linear strands that run opposite to
each other and twist together to form.
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. As a result
of semi-conservative replication, the
new helix will be composed of an
original DNA strand as well as a newly
synthesized strand. Cellular
proofreading and error-checking
mechanisms ensure near perfect fidelity
for DNA replication.
• In a cell, DNA replication begins at
specific locations, or
origins of replication in the genome
which contains the genetic material
of an organism. Unwinding of DNA
at the origin and synthesis of new
strands, accommodated by an
enzyme known as helicase, results
in replication forks growing bi-
directionally from the origin. A
number of proteins are associated
with the replication fork to help in
the initiation and continuation of
DNA synthesis. Most prominently,
DNA polymerase synthesizes the
new strands by adding nucleotides
that complement each (template)
strand. DNA replication occurs
during the S-stage of interphase.
Step 1: Replication Fork Formation
Before DNA can be replicated, the double
stranded molecule must be “unzipped” into two
single strands. DNA has four bases called
adenine (A), thymine (T), cytosine (C) and
guanine (G) that form pairs between the two
strands. Adenine only pairs with thymine and
cytosine only binds with guanine. In order to
unwind DNA, these interactions between base
pairs must be broken. This is performed by an
enzyme known as DNA helicase. DNA helicase
disrupts the hydrogen bonding between base
pairs to separate the strands into a Y shape
known as the replication fork. This area will be
the template for replication to begin.
• DNA is directional in both strands,
signified by a 5' and 3' end. This notation
signifies which side group is attached the
DNA backbone. The 5' end has a
phosphate (P) group attached, while the
3' end has a hydroxyl (OH) group
attached. This directionality is important
for replication as it only progresses in the
5' to 3' direction. However, the
replication fork is bi-directional; one
strand is oriented in the 3' to 5' direction
(leading strand) while the other is
oriented 5' to 3' (lagging strand). The
two sides are therefore replicated with
two different processes to accommodate
the directional difference.
Replication Begins : Step 2: Primer Binding

• The leading strand is the

simplest to replicate. Once
the DNA strands have been
separated, a short piece of
RNA called a primer binds
to the 3' end of the strand.
The primer always binds as
the starting point for
replication. Primers are
generated by the enzyme
DNA primase.
DNA Replication: Step 3: Elongation
• Enzymes known as DNA polymerases are responsible creating the new strand by a process
called elongation. DNA polymerase III binds to the strand at the site of the primer and begins
adding new base pairs complementary to the strand during replication. Because replication
proceeds in the 5' to 3' direction on the leading strand, the newly formed strand is continuous.
• The lagging strand begins replication by binding with multiple primers. Each primer is only
several bases apart. DNA polymerase then adds pieces of DNA, called Okazaki fragments, to
the strand between primers. This process of replication is discontinuous as the newly created
fragments are disjointed.