GENE DAMANGE AND REPAIR
Gene Damage
DNA Alteration and Repair
Transmission of genetic information
from one gen to anotheer requires:
- DNA to be repliated accurately
- Provision for repairing DNA
alterations rising sponteneously
and from exposure to DNA-
damaging environmenta
agents.
DNA Mutation
Beneficial = DNSA base-sequence
changes
- mutation provides genetic
variability that is raw material
of evolution.
Net rate at wc organisms
accumulate mutation = low
200,000 years for 1 mutation of an
average gene (but mutation rate
far greater)
DNA Damage repaired shortly after
it occurs so that future generations
will not be affected.
Never passed on to offsprings =
mutation occur in cells (other than
sperm and egg)
DNA Damage can occur
spontaneously or in response to
mutagens
Normal process = several mutation
occur simultaneously.
Spontaneous mutations
- depurination
- deamination
reactions are spontaneous hydrolysis
reactions
random interactions between DNA and
h20 molecules around it.
Depurination
loss of purine based by spontanous
hydrolysis of the glycosidic bond that
links it to deoxyribose
Deamination
Removal of base amino grou NH2
Can involve cytosine, adenine or
gunanine, changes the base-pairing
properties of affected base.
(cytosine most susceptible Uracil
(result)
Depurination and Deamination
hydrolytic reation
caused by random collision of water
molecules with bond that links the
amino group (for deamin.) of base to
the pyrimidine or purine ring.
Typical human cell: DNA Damage rate:
100 deaminations/day
Outcome of Spontaneous DNA
Mutation
* Erroneous base sequence may be
propagated when the strand serves as
a template in the next round of DNA
replication
Cytosine in deamination
* where a cytosine has been
converted to a uracil by deamination
the uracil behaves like thymine in its
base-pairing properties.
* It directs the insertion of an adenine
in the opposite strand, rather than
guanine, the correct base.
Effect: change in the amino acid
sequence and function of a protein
encoded by the affected gene.
Mutagens (Chemical or Radiation)
*Base analogs resemble nitrogenous
bases in structure and are
incorportated into DNA.
Base modifying agents (chemical
reaction) DNA bases structure
altered = DNA Adducts
DNA Adducts = chemical additional
products or intercalating agents
Intercalating agents insert
themselves between adjacent bases of
the double helix distorting DNA
structure
increasing the chance that a base
will be deleted or inserted during DNA
replication
Direct correlation = how much patient
smokes
Frequency of DNA
adducts
Endonucleases recruited to
DNA by proteins that recognizes
sites of DNA damage
-Repair endonucleases cleave DNA
backbone adjacent to damaged site
- other enzymes facilitates removal of
defective nucleotides
-DNA helicase might unwind the DNA
ocated between 2 nicks to release the
damaged SNA from double helix
-Alternatively, exonuclease might
attach to end created by a single nick
-exonuclease chew away the damaged
strand one nucleotide at a time

Radiation
Sunlight = strong source of UV
alters DNA triggering pyrimidine
dimer formation

How?
-Formation of covelant bonds between
adjecent pyrmindine bases = usually 2
thymines
- replication and transcription =
blocked by dimers
because enzyme carrying out these
fuctions cannot cope w the resulting
buldge in DNA double helix

Mutagens = radiation cont
Xrays or forms the emit radioactive
substances
= ionizing radiation
removes electrons from biological
molecules, thereby generating highly
reactive intermediates that cause
various types of DNA damage
Second
- missing nucleotides are replaced with
correct ones by DNA polymerase
- nucleotide sequence of the
complementary strand serves as a
template to ensure correct base
insertion, just as it does in DNA
replication.

GENE REPAIR
Purpose molecule important to Third
organisms health and survival hence -DNA ligase forms miessin
must maintain normal state. phosphoester bond = seals remaining
nick in repaired strand
Mechanisms:
First
-defective nucleotides cut out from one
strand of DNA double helix
carried out be Repair
Endonucleases

Excision Repair Pathways
* Base excision repair corrects singe
damanged bases in DNA
1 deamination
-detected by specific DNA glycosylases
recognize specific deaminated base
and remove it from the DNA molecule
( cleaving bond bet. Base and sugar
it is attached to)
2 depurination sugar with missing
base is recognized by a repair
endonuclease = detect depurination
- repair endonuclease nreaks
phosphodiester backbone on one side
of sugar lacking base
- second enzyme completes removal of
sugar-phosphate unit
* Nucleotide excision repair
removes pyrimidine dimers and other
bulky lesions in DNA
- utilize proteins that recognize major
distortion in DNA double helix
-recruits an enzyme = NER
endonuclease (excinuclease)
-makes two cuts in DNA
backbone, one on either side of
distortion.
-DNA helicase binds to stretch of DNA
bet. The nicks (29 nucleotides in
humans)
-unwinds it, freeing it from rest of DNA
-resulting gap filled in by DNA
polymerase and sealed by DNA ligase
NER System
-most versatile of a cells DNA repair
systems
-recognizing and correcting many
types of damage that cant otherwise
by repaired.
Some cases, NER system:
-specifically recruited to DNA regions
where transcription has been halted
because transcription machinery
encountered an area of DNA damage
(Transcription-coupled repair)
permits active genes to be repaired
faster than DNA sequences located
elsewhere in the genome.
Damaged NER
= results to individuals with disease
Xerodoma pigmentosum
(autosomal recessive disorder)
-carry a mutation in any of 7 genes
coding for components of NER system
-Result = cannot repair DNA damage
caused by UV rad in sunlight
=high risk of skin cancer
Mismatch Repair
Targets errord made during DNA
replication
improperly base-paired nucleotides
sometimes escape the normal
proofreading mechanisms
Mismatched bp dont hydrogen
bond properly presence can be
detected and correct by mismatch
repair system
Noncomplementary BP (Mismatch)
How can incorrect member of an
abnormal bp distinguished from
correct member? Pair is simply
composed of two normal bases
inappropraiely pair w each other
Mismatch and Repair
Mismatch
= the incorrect member of an AC base
pair were the base C and the repair
system instead removed the base A,
the repair system would create a
permanent mutation instead of
correcting a mismatched base pair!
Repair
=Recognize first which of the two DNA
strands was newly synthesized during
the previous round of DNA replication
(the new strand would be the one that
contains the incorrectly inserted base)
Mismatch Repair
1 Detection cells employs detection
system
2 Enzyme Rxn repair endonuclease
introduces single nick
3 Elimination exonuclease removes
incorrect nucleotides from nicked
strand.
Uracil and Thymine both paris with
adenine
Thymine has methyl group
= Because the methylation step used
to create thymine is energetically
expensive, it might seem more
efficient for DNA to contain uracil.
deamination damage repair
-DNA damaged through deamination
rxn, cytosine is converted to uracil
-detected and removes by DNA repair
enzyme = uracil-Dna glycosylase
If uracil is present in normal
component of DNA = dna repair
wouldnt work
because normal uracils would not be
distinguishable from uracils generated
by accidental deamination of cytosine
Using Thymine in DNA over uracil,
-cells ensure that DNA damage caused
by deamination of cytosine can be
recognized and repaired w no changes
in DNA molecule
Key Transition Points in Cell Cycle