Chapter 6 Pages 197-222

DNA Replication, Repair, and

Recombination

Copyright © Garland Science 2010

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 Objective

• Importance of Replication
• When Does Replication Occurs
• Mechanism of Replication
• Proteins that Mediate Replication
• Errors during replication
• How DNA is Damaged?
 Deficiency in the Removal of Old Cells
& Abrupt Cell Division Have Potential
to Cause Cancers Including Prostate Cancer
Figure 6-20 Essential Cell Biology (© Garland Science 2010)
Hereditary Information
is Passed
Faithfully
from “One Generation”
to the “Next”

Figure 6-1 Essential Cell Biology (© Garland Science 2010)

 Lymph
vessel
Tumor
Blood
vessel

Glandular Cancer
tissue cell
Metastatic
tumor
1 A tumor grows 2 Cancer 3 Cancer cells spread 4 Cancer cells
from a single cells invade through lymph and may survive
cancer cell. neighboring blood vessels to and establish
tissue. other parts of the a new tumor
body. in another part
of the body.
https://www.youtube.com/watch?v=vNXFk_d6y80
The ability of a cell to survive and proliferate in a chaotic
environment depends on the accurate duplication of the
vast quantity of genetic information carried in its DNA.

This is DNA replication.

The Significance of mutation and variation in Replication

DNA is duplicated at a very higher rate at 1000 nucleotides/sec

 DNA Replication
• Base pairing enables DNA replication.
• Each strand act as a template for the synthesis of
a new complementary strand.

Figure 6-3 Essential Cell Biology (© Garland Science 2010)

Three steps in DNA Replication

1. Initiation
2. Elongation
3. Termination
 Initiation
The positions at which the DNA is first opened are called replication origins.
A-T rich stretches of DNA are typically found at origin of replication.
These DNA sequences attract initiator proteins for replication.

Figure 6-5 Essential Cell Biology (© Garland Science 2010)

 Key Concepts
The Magic of complementarity and hydrogen bonds

In bacteria replication origins span 100 nucleotide pairs

A=T is weaker than G=C

Bacterial Genome is circular; it has one replication of origin

Human genome has 10000 overall and 220/chromosomes

Each replication origin has two forks that moves bidrectional

 Elongation
Replication forks move away in opposite direction from multiple replication
origins in eukaryotic cells. DNA molecules in the process of being replicated
contains Y-shaped junctions called replication fork.

Figure 6-9 Essential Cell Biology (© Garland Science 2010)

Figure 6-12 Essential Cell Biology (© Garland Science 2010)
a: template, b: leading strand, c: lagging strand, d: replication
fork, e: primer, f: Okazaki fragments
Figure 6-17a Essential Cell Biology (© Garland Science 2010)
 Enzyme Function in DNA replication
Also known as helix destabilizing enzyme. Unwinds the DNA double helix at
DNA Helicase
the Replication Fork.
Builds a new duplex DNA strand by adding nucleotides in the 5' to 3' direction. Also
DNA
performs proof-reading and error correction. There exist many different types of DNA
Polymerase
Polymerase, each of which perform different functions in different types of cells.
A protein which prevents DNA polymerase III from dissociating from the DNA parent
DNA clamp
strand.
Single-Strand Bind to ssDNA and prevent the DNA double helix from re-annealing after DNA helicase
Binding (SSB) unwinds it, thus maintaining the strand separation, and facilitating the synthesis of the
Proteins nascent strand.
Topoisomerase Relaxes the DNA from its super-coiled nature.
DNA Gyrase Relieves strain of unwinding by DNA helicase; this is a specific type of topoisomerase
Re-anneals the semi-conservative strands and joins Okazaki Fragments of the lagging
DNA Ligase
strand.
Provides a starting point of RNA (or DNA) for DNA polymerase to begin synthesis of
Primase
the new DNA strand. Primer is 10 nucleotides long synthesized every 100 nucleotides
Lengthens telomeric DNA by adding repetitive nucleotide sequences to the ends
Telomerase of eukaryotic chromosomes. This allows germ cells and stem cells to avoid the
Hayflick limit on cell division.
1. https://www.youtube.com/watch?v=cDwJTLnGEyw
 Deoxynucleotide triphosphate is required for Energy

Figure 6-10 Essential Cell Biology (© Garland Science 2010)

 DNA Polymerase is Self Correcting/Editing/Proof reading

Figure 6-13 Essential Cell Biology (© Garland Science 2010)

Figure 6-14 Essential Cell Biology (© Garland Science 2010)
Figure 6-16 Essential Cell Biology (© Garland Science 2010)
• Eukaryotic chromosomal DNA molecules
– Have at their ends nucleotide sequences, called telomeres, that postpone the erosion
of genes near the ends of DNA molecules

1 µm
Telomeres allow the completion of DNA synthesis at the ends of
eukaryotic chromosome:
Bacterial chromosomal DNA is circular; eukarytic chromosomal
DNA is linear.
Telomere in human: GGGGTTA repeat, humdreds
Telomerase: RNA template bearing reverse transcriptase

5’ 3’
Telomerase adds
3’ additional repeats
5’

5’ 3’
3’
5’

5’ 3’
3’
Replicated chromosome
 Telomerase Function

https://www.youtube.com/watch?v=vtXrehpCPEE
3.Termination. Telomerase replicates the ends of
eukaryotic chromosomes. :

Figure 6-18 Essential Cell Biology (© Garland Science 2010)

 A mismatch repair system removes replication errors that escape
the replication machine

Table 6-1 Essential Cell Biology (© Garland Science 2010)

Depurination of Guanine
Deamination of Cytosine
Thymine Dimer by Ultraviolet Light
Cells Possess a Variety of Mechanisms
for Repairing DNA
• Removal of damaged nucleotides by cleaving the covalent bond
• A repair DNA polymerase binds to the 3’-OH end of the cut DNA strand
• Then, fills the Gap by Complementary bases.
Figure 6-21 Essential Cell Biology (© Garland Science 2010)
 Replication Overview
The leading strand will be continuous - made longer by DNA
polymerase III
The lagging strand will not be continuous - contains Okazaki
fragments
Need to backfill
DNA polymerase III attaches to the RNA primer
The RNA primer is laid down by a DNA primase - not a RNA
primase!!!
Requires single-strand binding proteins to stabilize the DNA
DNA polymerase I will chop out the RNA primer and replace it
with DNA
DNA ligase will connect the Okazaki fragments together
Comparison Between DNA POL I and III
During DNA Replication:
DNA polymerase 3 synthesize DNA from 5' to 3' end
on the leading and lagging strand ( but stops at the
RNA Primer ) and has exo nuclease activity from 3' to
5' end for proof reading ( kind of a reverse gear )

On the other hand, DNA Polymerase 1 works on

lagging strand where it degrades RNA primer and
replace it with DNA ( synthesis from 5' to 3' end ) and
in contrast with DNA polymerase 3,it has proof reading
acivity from 5' to 3' end ( in the same direction )
Details of Bacterial and and Eukaryotic DNA Polymerases

https://en.wikipedia.org/wiki/DNA_polymerase