DNA

REPLICATION
Dede Renovaldi, M.Sc.(Biomed)
Faculty of Medicine & Health
Universitas Muhammadiyah Jakarta
 DNA Replication

Purpose: cells need to make a copy of

DNA before dividing so each daughter
cell has a complete copy of genetic
information.
 Base-Pairing Enables DNA Replication
In the preceding chapter, we saw that each strand of the DNA double helix
contains a sequence of nucleotides that is exactly complementary to the
nucleotide sequence of its partner strand. Each strand can therefore act
as a template, or mold, for the synthesis of a new complementary strand
 Semi-Conservative Model

Replication of DNA
• base pairing allows each strand to
serve as a template for a new
strand
• new strand is 1/2 parent template &
1/2 new DNA
 Bonding in DNA hydrogen
bonds
5 3

covalent
phosphodiester
bonds

3
5
 DNA Synthesis Begins at Replication Origins
• Site where replication begins
1 in E. coli
1,000s in human
• Strands are separated to
allow replication machinery
contact with the DNA
• Characterized by many A-T base
pairs because easier to break 2 H-
bonds that 3 H-bonds
• Note anti-parallel chains
 Anti-parallel strands
• Nucleotides in DNA backbone are
bonded from phosphate to sugar
between 5 & 3 carbons
• DNA molecule has “direction”
• DNA is synthesized in the 5’-to-3’
direction
• complementary strand runs in
opposite direction
New DNA Synthesis Occurs at Replication Forks
 The Replication Fork Is Asymmetrical

• The 5’-to-3’ direction of the DNA polymerization

mechanism poses a problem at the replication
fork.
• The sugar– phosphate backbone of each strand
of a DNA double helix has a unique chemical
direction, or polarity, determined by the way
each sugar residue is linked to the next, and
that the two strands in the double helix run in
opposite orientations.
• As a consequence, at the replication fork, one
new DNA strand is being made on a template
that runs in one direction (3’ to 5’), whereas the
other new strand is being made on a template
that runs in the opposite direction (5’ to 3’)
 How is DNA Synthesized?

• Simple addition of nucleotides along one strand, as

expected
 Called the leading strand
 DNA polymerase reads 3’  5’ along the leading
strand from the RNA primer
 Synthesis proceeds 5’  3’ with respect to the new
daughter strand
• Remember how the nucleotides are added!!!!! 5’  3’
 How is DNA Synthesized?

• Other daughter strand is also synthesized 5’3’ because

that is only way that DNA can be assembled.
• However the template is also being read 5’3’
• Compensate for this by feeding the DNA strand through
the polymerase, and primers and make many short
segments that are later joined (ligated) together
• Called the lagging strand
• Those short fragments are called as Okazaki Fragments
DNA Replication
Large team of enzymes coordinates replication
Replication
Steps
 -Unwind DNA-
• Helicase enzyme Replication: 1st step
unwinds part of DNA helix
and stabilized by single-stranded binding proteins
= PREVENTS DNA MOLECULE FROM CLOSING!
• DNA gyrase
Enzyme that prevents tangling upstream from the replication fork
helicase gyrase

single-stranded binding
proteins
replication fork
RNA Primase
Replication: 2nd step
• Adds small section of RNA to act as primers to the 3’ end of
template DNA
Why must this be done?
• DNA polymerase 3 (enzyme that builds new DNA strand) can
only add nucleotides to existing strands of DNA
 Replication: 3rd step

 Build daughter DNA

strand
 With the help of the enzyme
DNA polymerase III

DNA
Polymerase III
 Replication: 4th step

• Replacement of RNA primer by DNA (Removal of Primers)

• Done by DNA polymerase I
• Other enzymes needed to excise (remove) the primers
 Nuclease – removes the RNA primer nucleotide by
nucleotide
 Repair polymerase – replaces RNA with DNA
 DNA ligase – seals the sugar-phosphate backbone by
creating phosphodiester bond
 Requires Mg2+ and ATP
Proteins at a Replication Fork Cooperate to Form a
Replication Machine
 Other Necessary Proteins

• Helicase opens double helix and helps it

uncoil
• Single-strand binding proteins (SSBP) keep
strands separated – large amount of this
protein required
• Sliding clamp
 Subunit of polymerase
 Helps polymerase slide along strand
• All are coordinated with one another to
produce the growing DNA strand (protein
machine)
 Mistakes during Replication

• Base pairing rules must be maintained

 Mistake = genome mutation, may have
consequence on daughter cells
• Only correct pairings fit in the DNA
polymerase active site
• If wrong nucleotide is included
 DNA Polymerase uses its proofreading
ability to cleave the phosphodiester bond
of improper nucleotide
 Activity 3’  5’
 And then adds correct nucleotide and
proceeds down the chain again in the 5’ 
3’ direction
Replication of DNA
DNA
polymerase III lagging strand
DNA
polymerase I
3’
Okazaki primase
fragments 5’
5’ ligase
3’ 5’ SSB

3’ helicase

DNA
polymerase III
5’ leading strand
3’
direction of replication
SSB = single-stranded binding proteins
 How about telomere?

A telomere is the end of a chromosome. Telomeres are made of

repetitive sequences of non-coding DNA that protect the chromosome
from damage. Each time a cell divides, the telomeres become shorter.
Eventually, the telomeres become so short that the cell can no longer
divide.

Telomerase replicates the ends of

eucaryotic chromosomes
References
 THANKS
Do you have any questions?
Find me at :
dede.renovaldi@gmail.com

Wanna watch an action movie?

Here!
https://www.youtube.com/watch?v=TN
KWgcFPHqw&ab_channel=yourgenome