DNA repair

spontaneous vs. induced mutation

gametic vs. somatic mutation
lethal or conditional mutation
Classification of the mutation
Spontaneous mutations
are those that happen naturally
no specific agents are associated with their
occurence
and they are generally assumed to be random
changes in the nucleotide sequences of genes
Induced mutations
those that result from the influence of any
artificial factor
various forms of radiation
a wide spectrum of chemical agents
biological agents (e.g. viruses)
Gametic vs. somatic mutations
Mutation arising in somatic cells are not
transmitted to future generations
Mutations in gametes or gamete-forming tissue
are of greater significance because they are
transmitted to offspring as part of the germ line
dominant
recessive
X-linked
Lethal vs. conditional mutations
Mutation may interrupt a process that is essential
to the survival of the organism
- in this case, it is reffered to as a lethal mutation
Conditional mutation is present in the genome of
an organism, but it is expressed and can be
detected only under certain conditions
Induced Induced mutagenesis mutagenesis
Can be caused by environmentaI agents
that damage DNA:
UV Iight
X- rays and -rays
ChemicaI carcinogens e.g. cigarette smoke
DNA damage can Iead to mutations unIess it
is removed by DNA repair enzymes
Unrepaired damage can have serious
consequences
IMPORTANCE OF DNA REPAIR
Hoeifmakers, 2001
Photoactivation Repair in E. coIi
Exposing UV treated ceIIs to bIue Iight
resuIts in a reversaI of the thymine dimer
formation
Enzyme, photoactivation repair enzyme
(PRE) absorbs a photon of Iight (from bIue
Iight) and is abIe to cIeave the bond
forming the thymine dimer.
Once bond is cIeaved, DNA is back to
normaI
Excision Excision Repair Repair
Conserved throughout evoIution, found in
aII prokaryotic and eukaryotic organisms
Three step process:
- 1. Error is recognized and enzymaticaIIy cIipped out
by a nucIease that cIeaves the phosphodiester bonds
(uvr gene products operate at this step)
- 2. DNA PoIymerase I fiIIs in the gap by inserting the
appropriate nucIeotides
- 3. DNA Ligase seaIs the gap
Excision Excision Repair Repair
Two know types of excision repair
- Base Base excision excision repair repair (BER) (BER)
corrects damage to nitrogenous bases created by
the spontaneous hydroIysis of DNA bases as weII
as the hydroIysis of DNA bases caused by agents
that chemicaIIy aIter them
- NucIeotide NucIeotide excision excision repair repair (NER) (NER)
Repairs "buIky" Iesions in DNA that aIter or distort
the reguIar DNA doubIe heIix
Group of genes (uvr) invoIved in recognizing and
cIipping out the Iesions in the DNA
Repair is compIeted by DNA poI I and DNA Iigase
Proofreading Proofreading and and Mismatch Mismatch Repair Repair
In bacteriaI systems, proofreading proofreading decreases the
error rate in DNA repIication by two orders of
magnitude
- from 1 mismatch in every 105 nucIeotide pairs to 1
in every 107 base pairs
Mismatch Mismatch repair repair is another mechanism by which
mismatches can be fixed in the DNA strand
In bacteria, mismatch repair is based on the
process of DNA DNA MethyIation MethyIation, which IabeIs one
strand, providing a basis for the mismatch repair.
Post Post- -RepIication RepIication Repair Repair
Post-replication repair Discovered in
by Miroslav Radman
Responds when damaged DNA escapes
repair and the damage disrupts replication
Rec A protein stimulates recombination
between donor strand and new strand
Creates gap in donor strand which can
be repaired
DNA Polymerase and DNA Ligase involved
Xeroderma pigmentosum (XP): Patients are
hypersensitive to UV light; patients often develop
malignancies of the skin.
Ataxia teIangiectasia (AT): Patients are sensitive
to gamma irradiation; patients develop
neurological and skin lesions.
Fanconi's anemia: Patients demonstrate aplastic
anemia, growth retardation, and congenital
anomalies; related to a deficiency in repair of
DNA cross-links.
Diseases in which DNA repair is damage
Xeroderma Pigmentosum (XP) and DNA
Repair Defects
XP is an autosomaI recessive disease associated
with dry skin, freckIing, corneaI uIceration, and skin
tumors
Many patients die before age 30 from metastases
of maIignant skin tumors
One form of XP is produced by a defect in the
human endonucIease that removes pyrimidine
dimers
Mutations in at Ieast seven other genes invoIved
in repairing UV-damaged DNA can cause XP
DNA Repair and Clinical Syndromes:
Increased Sensitivity;
Chromosomal Instability
and Increased Cancer Risk
Xeroderma Pigmentosum (XP)
Symptoms include:

Extreme sensitivity
to sunlight

Early onset of skin

cancer
XP cases
SunIight-induced dermatoIogic abnormaIities in a
patient with xeroderma pigmentosum.
TypicaI skin manifestation of
xeroderma pigmentosum with
numerous areas of hypopigmentation
and freckIes (ie, soIar Ientigines) with
different intensities of pigmentation.
NER
Healthy
DNA
Why are XP patients sensitive to
sunIight?
HeaIthy
DNA
UV
Damaged
DNA

NER
Healthy
DNA
Why are XP patients sensitive to
sunIight?
HeaIthy
DNA
UV
Damaged
DNA
NO
REPAIR

XP XP: Skin Cancer Incidence Rapid : Skin Cancer Incidence Rapid

Secondary to NER Faulty Repair of UV Secondary to NER Faulty Repair of UV- -induced induced
DNA Damage and Genetic NER Mutations DNA Damage and Genetic NER Mutations
MoIecuIar Genetics of Xeroderma Pigmentosum
Error prone DNA
photoproduct bypass
poIymerase
6p21.1-p12 POLH XP variant
DNA-repair protein
compIementing XP-G
ceIIs
13q33 ERCC5 XPG G
DNA-repair protein
compIementing XP-F
ceIIs
16p13.3-p13.13 ERCC4 XPF F
DNA damage binding
protein 2
11p12-p11 DDB2 XPE E
TFIIH basaI
transcription factor
compIex heIicase
subunit
19q13.2-q13.3 ERCC2 XPD D
DNA-repair protein
compIementing XP-C
ceIIs
3p25 XPC XPC C
TFIIH basaI
transcription factor
compIex heIicase XPB
subunit
2q21 ERCC3 XPB B
DNA-repair protein
compIementing XP-A
ceIIs
9q22.3 XPA XPA A
Protein Name ChromosomaI Locus Gene SymboI Locus Name
CompIementation
Group
Data are compiled from the following standard references: Gene symbol from HUGO; chromosomal locus, locus name, critical region,
complementation group from OMM; protein name from Swiss-Prot.
Fanconi anemia (FA)
Fanconi anemia (FA) is an autosomaI recessive
disease characterized by progressive bone marrow
faiIure due to defective stem ceII function.
FA ceIIs are hypersensitive to DNA cross-Iinking
agents such as mitomycin C (MMC) resuIting in
cytogenetic aberrations, G2-M ceII cycIe arrest,
apoptosis, and ceII death.
Seven compIementation groups (termed FANCA-G)
are identified. Group A (FANCA) mutations
are the most prevaIent (70%).

The function of the FANC genes are stiII uncIear.

InvoIvement in DNA repair system is suggested.
There are at least seven FA genes: A, C, D2, E, F, G and
BRCA2

Cockayne's Cockayne's Syndrome Syndrome

Occurrence: 1 per miIIion popuIation
Sensitivity: uItravioIet radiation (sunIight)
Disorder: arrested deveIopment, mentaI retardation,
dwarfism, deafness, optic atrophy, intracraniaI
caIcifications; (no increased risk of cancer)
BiochemicaI: defect in NER
Genetic: autosomaI recessive, five genes (A, B
and XPB, D & G)
Cockayne's Syndrome
Trichothiodystrophy Trichothiodystrophy
Occurrence: 1-2 per miIIion popuIation
Sensitivity: uItravioIet radiation (sunIight) in
subset of patients
Disorder: suIfur deficient brittIe hair, mentaI and
growth retardation, pecuIiar face with receding
chin, ichthyosis; (no increased cancer risk)
BiochemicaI: defect in NER
Genetic: autosomaI recessive, three genes (TTDA,
XPB, XPD)
Trichothiodystrophy
Ataxia teIangiectasia (AT):