Mechanism of Rolling Circle

replication
 Introduction
• Rolling circle replication is a process of
unidirectional nucleic acid replication.
• It can rapidly synthesize multiple copies of
circular molecules of DNA or RNA, such as
plasmid.
• Eukaryotic also replicate their DNA/RNA
via this methods.
• Widely used in molecular biology &
biochemical nanotechnology, especially in
the field of biosensing (as a method of
signal amplification).
 DNA rolling circle replication has five Steps

• Circular ds DNA will be nicked

• 3’ end is elongated using un nicked DNA as
leading strand (template)
• 5’ end displaced is as a lagging strand and is
made double stranded DNA via a series of
OKAZAKI fragments
• Replication of both un nicked and displaced
ss DNA
• Displaced DNA circulates and synthesis its
own complementary strand
 Three steps of Rolling Circle Replication
(1) Initiation
• In the process of initiation, three enzymes works
together such as:
1. Helicase
2. Topoisomerase
3. Single stranded binding proteins (SSBPs)
• These enzymes bind with phosphate ends of nick
strand
(2) Elongation
• In the process of elongation, 3’OH group of broken
strand, work as a template
• The polymerase enzyme help to move in a circle for
elongation, due to which it is named as rolling circle
mechanism
• 5’ end will be displaced and grow like as a thread
(3)Termination
• At the point of termination, the linear DNA molecule is
cleaved from the circle resulting in a double stranded
circular DNA molecule and a single stranded DNA
molecule
• The linear single stranded molecule is circularized by
the action of ligase and then replication to double
stranded circular plasmid molecule
Replication starts at ORI region
by a initiator Rep A

Region bind with DNA polymerase and

Replication starts
 Progeny positive strand
Helicase also attach to the
synthesis
region
Positive strand also synthesis its
complementary strand
 Breakage is joining by
ligase

Progeny strand can also synthesize another

strand of DNA
 Examples

Viral DNA: Some DNA viruses replicate their genomic

information in host cells via rolling circle replication

• Human herpes virus

• Human papilloma virus

• Gemini virus

Viral RNA: Some RNA viruses and viroids also replicate

their genome through rolling circle RNA replication

• Pospiviridiae family

• Avsunviridiae family
Thank you