DNA Replication in

Prokaryotes
GROUP 1- ASIS, ANGWANI, CLOR, &
CUMAMAO
DNA Replication

• DNA replication is a
biological process which
produces two identical
daughter DNA strands from
a double stranded parental
DNA.
• A basis for biological
inheritance in all living
organisms.
Prokaryotic DNA Replication

• DNA replication is semi

conservative.

• DNA replication is
bidirectional.

• DNA replication is semi

discontinuous.
Replicon

• A DNA segment that undergoes

replication.
• It consists of an origin where replication
begins and a terminus where replication
stops.
• E. coli has a single replicon on its
chromosome, as do most prokaryotes.
Origin of Replication
• Ori C is initiated at the origin of replication
• This sequence of bps is recognized by a protein
that binds to this cite and opens up the DNA.
• Helicase unwinds the DNA
• ATP hydrolysis is required for this process
 Bidirectional Replication
• As the DNA opens up, two Y-
shaped structures called
replication forks are formed at
the origin of replication making a
replication bubble.
• The replication forks get extended
in opposite directions as the
replication proceeds.
• Single stranded binding proteins
coat the single strand of DNA near
the replication fork to prevent
single stranded DNA from winding
back into double helix.
DNA Polymerases
• DNA polymerases refers to a group of enzymes
that is responsible for DNA synthesis.
Prokaryotic Polymerases

• Three main types of Polymerases:

Leading Strand
• as we all know DNA is antiparallel
• the polymerases can only extend in 5' to 3'
direction which poses a slight problem at
replication fork.
• one strand (5' to 3' ) which is complementary to 3'
to 5' parental DNA strand is synthesized
continuously towards the replication fork
• this continuously strand is known as leading
strand
Lagging Strand
• the other strand complementary to 5' to 3' parental DNA
is extended away from the replication fork in small
segments (Okazaki fragments), each requiring a primer
to start synthesis.
• the strand with Okazaki fragments ( named after the
scientist who discovered it) is known as lagging strand
Enzymes and Proteins involved in Replication

• Helicase
• Single strand
binding proteins
• topoisomerase
• Primase
• DNA polymerase III
• DNA polymerase I
• DNA ligase
Three main steps in the process of DNA Replication:
Initiation
• Start or to initiate something.
• During prokaryotic DNA replication the protein DnaA
(chromosomal replication initiator protein) bind to the origin of
replication while DnaB helicase unwinds the DNA helix and
two replication forks are formed at the origin of replication.
• It is the bidirectional replication.
The main events involved in initiation:

• Recognition of origin.
•DNA melting.
•Stabilization of single strand.
•Assembly of Primosome at the two forks
produced.
•Start synthesis of two daughter strands.
Proteins for initiation
• Replication initiation in E.coli
requires 6 proteins.
• DnaA or chromosomal
replication initiator protein
• DnaB or replicative DNA
helicase
• DnaC (DNA replication protein)
• SSBp (single stranded DNA-
binding protein)
• DNA gyrase
• DnaG primase
Elongation
• The synthesis of two new daughter strands
takes place complementary to the template
strands.
• DNA polymerases are the enzymes that
synthesize the new daughter DNA molecules.
• Three types of DNA polymerases in prokaryotes
are:
DNA polymerase I
DNA polymerase II
DNA polymerase III
• DNA polymerase I- Enzyme that has exonuclease
activity in which it removes RNA primer. Replace it
DNA sequences.
• DNA polymerase II- Enzymes that catalyzes the repair
of nucleotide bas pairs.
• DNA polymerase III- Enzyme that synthesizes the
daughter strands and also adds nucleotides one by
one to the growing DNA chain. Main enzyme that adds
in the 5ʹ to 3ʹ direction. DNA polymerase III uses 3ʹ-
hydroxyl group of the RNA primer as an acceptor of the
first DNA sequence.
B subunit enzyme

Proofreading

Ligase
 Termination

• The ending of transcription,

and occurs when RNA
polymerase crosses a stop
(termination) sequence in
the gene.
• Release factor recognizes
stop codon, translational
complex dissociates, and
completed polypeptide is
release.
• E.Coli DNA. Termination sites
like A, B, C, D, F and G are
found to present in DNA. Of
these sites, Ter A terminates
the counter clockwise moving
fork while ter C terminates the
clockwise moving forks.

• Tus protein binds with Ter to

form a Tus-Ter complex.
• Ter sequence have two
binding sites permissive and
Non-permissive.
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