DNA Replication Process in Prokaryotes

It is the process by which a prokaryote repeats its DNA into another copy that is passed
on to daughter cells which is known as prokaryotic DNA replication (PDR).

Prokaryotes have been extensively investigated in terms of DNA replication, partly due to the tiny
size of their genomes and the large number of mutations that are available. 4.6 million base pairs
make up a circular chromosome in the bacteria E. coli, and the entire chromosome is duplicated in
around 42 minutes, starting from a single origin of replication and progressing around the circle both
ways. Approximately 1000 nucleotides are added every second, according to this calculation. The
technique is relatively quick and does not involve many mistakes.

During the process of DNA replication, a great number of proteins and enzymes are involved, each
of which plays an important function in the process. In particular, the enzyme DNA polymerase (also
known as DNA pol), which adds nucleotides one at a time to the expanding DNA chain that are
complementary to the template strand, serves as an important cog in the chain-building machine.
Adding nucleotides requires energy, which is obtained from nucleotides that have three phosphates
linked to them, similar to how ATP has three phosphate groups attached. Nucleotides that have
three phosphates attached to them are known as triphosphate nucleotides. It takes energy to break
down the link between the phosphates, and that energy is then channelled towards the formation of
the phosphodiester bond between the incoming nucleotide and the expanding chain. DNA
polymerases are known to exist in three different forms in prokaryotes: DNA pol I, DNA pol II, and
DNA pol III. In recent years, scientists have discovered that DNA pol III is the enzyme responsible
for synthesis, while DNA pol I and DNA pol II are predominantly responsible for repair.

DNA Replication Steps

The following are the critical phases in the replication of genetic material:

Initiation
In order to ensure that DNA replication is successful, it must be performed with extreme precision
because even the smallest error could result in mutations. As a result, DNA replication cannot begin
at any location in the genome at random.

The origin of replication is a specific location that must be reached in order for replication to occur.
This is the stage at which the replication process begins to unfold. The marking of this origin is
followed by the unwinding of the two DNA strands, which marks the beginning of replication.
 Due to the large amount of energy required, unzipping DNA strands throughout their whole length is
not practical. The replication fork is formed initially, and the DNA strand is unzipped by the helicase
enzyme, which catalyses the replication fork formation.

Elongation
In parallel with the separation of the strands of DNA, the polymerase enzymes begin synthesising
the corresponding sequence in each strand of DNA that has been split. The parental strands will
serve as a template for the synthesis of daughter strands that are produced in the future.

In addition, it should be emphasised that elongation is unidirectional, meaning that DNA is always
polymerised in just one direction, from 5′ to 3′. As a result, on one strand (the template 3’5′),
replication is continuous, therefore the term “continuous replication,” whereas on the other strand
(the template 5’3′), replication is discontinuous, so the term “discontinuous replication.” They are
found in the form of pieces known as Okazaki fragments. Later on, an enzyme known as DNA ligase
links them together.

Termination
The termination of replication occurs in a variety of ways depending on which organism is being
considered. Chromosomes in organisms like E.coli are round in shape. The same thing occurs when
the two replication forks between the two terminals come into contact with each other.

Steps of DNA Replication In prokaryotes

 The DNA replication process is bidirectional, and it begins at a point on the DNA molecule known as
the origin of replication, where it continues in both directions.

 It is at this point that enzymes unravel the double helix structure of DNA, allowing its constituent
parts to be replicated.

 In order for a pair of replication forks to develop, the helix must be unwound by the helicase enzyme,
and the unwound helix must be stabilised by SSB proteins and DNA isomerases.

 Primase synthesises RNA primers with a base sequence of 10 bases, which are used to trigger the
synthesis of both the leading and lagging strands.
 In the 5′ to 3′ direction, DNAP III (DNA Polymerase III) continues to synthesise the leading strand. In
the 5′ to 3′ direction, DNAP III (DNA Polymerase III) continues to synthesise the lagging strand, but
the synthesis is terminated by the creation of Okazaki fragments.

 DNA polymerase is a type of enzyme. Removes the RNA primers with a base length of 10 bases
and fills in the gap with deoxynucleotides.

 Then, using DNA ligase, the breaks between Okazaki fragments as well as the gaps around the
primers are sealed, resulting in continuous strands.

 The cytoplasm of the cell is where the entire process of replication takes place.
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