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DNA_replication

DNA replication is the process of duplicating the entire genome before cell division, requiring extreme accuracy to maintain genomic integrity. The replication is semi-conservative, occurring in the 5'-3' direction, and involves various proteins such as DNA polymerase and helicases to facilitate the process. Eukaryotes have multiple origins of replication, and the mechanism includes initiation, elongation, and termination phases.

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Sunil Tiwari
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3 views

DNA_replication

DNA replication is the process of duplicating the entire genome before cell division, requiring extreme accuracy to maintain genomic integrity. The replication is semi-conservative, occurring in the 5'-3' direction, and involves various proteins such as DNA polymerase and helicases to facilitate the process. Eukaryotes have multiple origins of replication, and the mechanism includes initiation, elongation, and termination phases.

Uploaded by

Sunil Tiwari
Copyright
© © All Rights Reserved
Available Formats
Download as PDF, TXT or read online on Scribd
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DNA replication

DNA stores information in the sequence of


its bases
Much of DNA’s sequence-specific information is accessible only
when the double helix is unwound

Proteins read the DNA sequence of nucleotides as the DNA helix


unwinds.

Proteins can either bind to a DNA sequence, or initiate the copying


of it.

Ø Some genetic information is accessible even in intact, double-


stranded DNA molecules

Ø Some proteins recognize the base sequence of DNA without


unwinding it (One example is a restriction enzyme).
DNA Replication
Process of duplication of the entire genome prior to cell
division

Biological significance
n extreme accuracy of DNA replication is necessary in
order to preserve the integrity of the genome in
successive generations
n In eukaryotes , replication only occurs during the S
phase of the cell cycle.
n Replication rate in eukaryotes is slower resulting in a
higher fidelity/accuracy of replication in eukaryotes
Basic rules of replication
A. Semi-conservative
B. Starts at the ‘origin’
C. Synthesis always in the 5-3’ direction
D. Can be uni or bidirectional
E. Semi-discontinuous
F. RNA primers required
DNA replication
3 possible
models
Semi-conservative
replication:
One strand of
duplex passed on
unchanged to each
of the daughter
cells. This
'conserved' strand
acts as a template
for the synthesis of
a new,
complementary
strand by the
enzyme DNA
polymerase
B) Starts at origin
Initiator proteins identify specific base
sequences on DNA called sites of origin

Prokaryotes – single origin site E.g E.coli - oriC


Eukaryotes – multiple sites of origin (replicator)
E.g. yeast - ARS (autonomously replicating
sequences)

Prokaryotes Eukaryotes
In what direction does DNA replication occur?
C) Synthesis is ALWAYS in the 5’-3’ direction
What happens if a base
mismatch occurs? Where does energy for addition
of nucleotide come from?
Why does DNA replication only occur in the 5’ to 3’ direction?

Should be PPP here


D) Uni or bidirectional
n Replication forks move in one or opposite directions
E) Semi-discontinuous replication
Anti parallel strands replicated simultaneously
q Leading strand synthesis continuously in 5’– 3’

q Lagging strand synthesis in fragments in 5’-3’


Semi-discontinuous replication
New strand synthesis always in the 5’-3’ direction
F) RNA primers required
Core proteins at the replication fork
Topoisomerases - Prevents torsion by DNA breaks
Helicases - separates 2 strands
Primase - RNA primer synthesis
Single strand - prevent reannealing
binding proteins of single strands
DNA polymerase - synthesis of new strand
Tethering protein - stabilises polymerase
DNA ligase - seals nick via phosphodiester
linkage
The mechanism of DNA replication
n Initiation
n Proteins bind to DNA and open up double helix
n Prepare DNA for complementary base pairing
n Elongation
n Proteins connect the correct sequences of
nucleotides into a continuous new strand of DNA
n Termination
n Proteins release the replication complex
The mechanism of DNA replication

http://www.thelifewire.com
Life: 7th ed - Chapter 11
Core proteins at the replication fork

Nature (2003) vol 421,pp431-435 Figure in ‘Big’ Alberts too


Further Reading material:
https://engineering.ucsb.edu/~shell/che170/DNA-notes.pdf

What kind of enzyme synthesizes the


new DNA strand?
1) RNA polymerase

2) DNA Polymerase

3) Primase

4) Helicase

5) Topoisomerase
Eukaryotic chromosomes have
multiple origins of replication
1. True
2. False
In what direction is the newly
synthesized DNA produced?
1. 5'-3'

2. 3'-5'

3. In the direction of the major groove

4. Both 5'-3' and 3'-5' depending on which


strand is being replicated
Nucleotides are always added to the growing DNA
strand at the 3’ end, at which the DNA has a free
______ on the 3’ carbon of its terminal deoxyribose.

1. Phosphate group
2. Hydroxyl group
3. Nitrogen base

4. Methyl group
The E. coli chromosome has 4.7x106 bp;
a bi-directional replication fork
progresses at about 1000
nucleotides/sec. Therefore, the minimum
time required to complete replication is

1) 12 min.
2) 24 min.
3) 39 min
4) 78 min
5) 120 min
What is the sequence (1 to 6) in which
these proteins function during DNA
replication

n ____ RNA primase


n ____ DNA ligase
n ____ DNA polymerase
n ____ Topoisomerase
n ____ DNA helicase
n ____ tethering proteins
Why is an RNA primer necessary for DNA
replication?
A. The RNA primer is necessary for the activity of
DNA ligase.

B. The RNA primer creates the 5’ and 3’ ends of the


strand.

C. DNA polymerase can only add nucleotides to RNA


molecules.

D. DNA polymerase can only add nucleotides to an


existing strand

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