DNA Repair Mechanisms-1 PDF

DNA Repair Mechanisms
Living organisms contain many enzymes that scan

their DNA for damage and initiate repair processes
when damage is detected.
1
DNA Repair Mechanisms in E. coli
1. Light-dependent repair (photo-reactivation)

2. Excision repair
3. Mismatch repair
4. Post-replication repair
5. Error-prone repair system (SOS response)
2
1) UV Light-Dependent
Repair: Photolyase
Cleaves Thymine
Dimers.
--No Endonuclease
--No Ligase
3
2) Types of Excision Repair
• Base excision repair pathways remove
abnormal or chemically modified bases.
• Nucleotide excision repair pathways remove

larger defects, such as thymine dimers.
4
Excision Repair
• A DNA repair endonuclease or endonuclease-
containing complex recognizes, binds to, and
excised the damaged base or bases.
• A DNA Polymerase fills in the gap, using the

undamaged complementary strand of DNA as a
template.
• DNA ligase seals the break left by DNA

polymerase.
5
Base Excision Repair
AP:apyrimidinic site
6
( )
7
Nucleotide Excision Repair
8
9
3) Mismatch Repair in E. coli
• The mismatched nucleotide is excised from the new
strand and replaced with the correct nucleotide, using the
methylated parental strand as a template.
• Mismatching or mispairing of G and T
• The A in GATC sequences is methylated subsequent to

DNA replication.
• In newly replicated DNA, the parental strand is

methylated, but the new strand is not. This difference
allows the mismatch repair system to distinguish the new
strand from the old strand.
10
• MutS recognizes
mismatches and binds to
them to initiate the repair
process.
• MutH and MutL join the
complex.
• MutH cleaves the
unmethylated strand at
hemimethylated GATC
sequences on either side of
the mismatch.
• Excision requires MutS,
MutL, MutU (DNA helicase
II), and an exonuclease.
11
• DNA polymerase III
fills in the gap, and
DNA ligase seals
the nick.
12
4) Post-replication Repair in E. coli
• A thymine dimer in the template strand blocks replication
(DNA Polymerase III does not recognize thymidine dimer)
• DNA Polymerase III restarts DNA synthesis past the dimer,

leaving a gap in the nascent strand.
• RecA binds to the single strand of DNA at the gap and

mediates base pairing with the homologous segment of
the sister double helix to fill the gap.
• DNA polymerase fills the gap in the sister double helix,

and DNA ligase seals the nick.
13
14
5) The SOS Response in E. coli
• If DNA is heavily damaged by mutagenic agents, the SOS

response, which involves many DNA recombination, DNA
repair, and DNA replication proteins, is activated.
• DNA dependent DNA Polymerase V replicates DNA in

damaged regions, but sequences in damaged regions cannot
be replicated accurately.
• This error-prone system eliminates gaps but increases the

frequency of replication errors (Pol II, IV and V are low-
fidelity polymerases)
15
Induction of the SOS Response
• In the absence of DNA damage, LexA binds to DNA
regions that regulate transcription of SOS response genes
and keeps their expression levels low.
• When extensive DNA damage occurs, RecA binds to
single-stranded regions of DNA in damaged regions.
• This activates RecA, which stimulates LexA to inactivate
itself. When LexA is inactivated, the SOS response genes
are expressed.
16
Human Diseases with Defects in
DNA Repair
Several human disorders result from defects in
DNA repair pathways.
17
Xeroderma Pigmentosum (XP)
• Individuals with XP are sensitive

to sunlight (UV light).
• The cells of individuals with XP are

deficient in the repair of UV-
induced damage to DNA.
• Individuals with XP may develop

skin cancer or neurological
abnormalities.
18
Mutations in Human Globin Genes
• Adult hemoglobin (Hemoglobin A) contains two 
chains and two  chains.
• Hemoglobin in patients with sickle-cell anemia

(Hemoglobin S) differs from Hemoglobin A at only
one position.
• The sixth amino acid in the  chain is glutamic acid in

Hemoglobin A (HBBA) and is valine in Hemoglobin S
(HBBS). This substitution is caused by mutation of a
single base pair (T:A substitution).
19
Tay-Sachs Disease
• Tay-Sachs disease is an
autosomal recessive
disease.
• The mutation causing
Tay-Sachs disease is in
the gene encoding
hexosaminidase A.
not linked to sex chromosome

20
 Transgenic Mouse: Generic term for an engineered mouse that has a
normal DNA sequence for a gene replaced by an engineered sequence or a
sequence from another organism.
 Knockout Mouse: A transgenic mouse in which the normal gene is missing

or engineered so that is not transcribed or translated. “Knocks out” that
gene.
 Knockin Mouse: A transgenic mouse in which the engineered “transgene”

is subtly manipulated to: (A) alter the function of the gene (e.g., replace one
amino acid with another in a site to determine if that site is essential for the
protein’s function); (B) change transcription rate to overproduce or
underproduce the gene product; or (C) create a fluorescent gene product to
map its distribution in tissue.
 Conditional Knockout (Knockin) Mouse: A transgenic mouse in which

the transgene is knocked out (or in) in specific tissues, at a specific
developmental stage, or in response to an exogenous substance (e.g., an
antibiotic). 21

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DNA Repair Mechanisms

Living organisms contain many enzymes that scan

1. Light-dependent repair (photo-reactivation)

• Nucleotide excision repair pathways remove

• A DNA Polymerase fills in the gap, using the

• DNA ligase seals the break left by DNA

• Mismatching or mispairing of G and T

• The A in GATC sequences is methylated subsequent to

• In newly replicated DNA, the parental strand is

• DNA Polymerase III restarts DNA synthesis past the dimer,

• RecA binds to the single strand of DNA at the gap and

• DNA polymerase fills the gap in the sister double helix,

• If DNA is heavily damaged by mutagenic agents, the SOS

• DNA dependent DNA Polymerase V replicates DNA in

• This error-prone system eliminates gaps but increases the

• Individuals with XP are sensitive

• The cells of individuals with XP are

• Individuals with XP may develop

• Hemoglobin in patients with sickle-cell anemia

• The sixth amino acid in the  chain is glutamic acid in

not linked to sex chromosome

 Knockout Mouse: A transgenic mouse in which the normal gene is missing

 Knockin Mouse: A transgenic mouse in which the engineered “transgene”

 Conditional Knockout (Knockin) Mouse: A transgenic mouse in which

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