MUTATION &

REPAIR
SYSTEM
DESAK MADE WIHANDANI
DNA : THE MOLECULE OF LIFE
/REPLICATION
REPLICATION
 MUTATION
• Any change in the nucleotide (base) sequence of the genetic
material (DNA or RNA)
• Set in DNA after a second round of replication
• Can be detected by comparing the mutated organism (mutant)
with another nonmutated organism of the same type (wild type)
• The effect is depend on size (single to millions bases) and
location of mutation
 Mutation
• SPONTAN
replication error

• INDUCIBLE
mutagen (chemist)
ultraviolet dll
• Base Substitution (Point Mutation):
o Transition
(PurinePurine, PyrimidinePyrimidine)
o Transvertion
(Purine  Pyrimidine)

• Non Base Substitution :

o Deletion
o Insertion
 Point mutation

 Transition A G
purine replaces purine
or C T
pyrimidine replaces pyrimidine

 Transversion

purine replaces pyrimidine A G

or
pyrimidine replaces purine
C T
SUBSTITUTION

MISSENSE MUTATION
SUBSTITUTION

NON-SENSE MUTATION
 Nonsense mutation
A point mutation changing a codon
for an amino acid into a stop
codon (UAA, UAG or UGA).
5’ ATG GGA GCT CTA TTA ACC TAA 3’
met gly ala leu leu thr stop

5’ ATG GGA GCT CTA TGA ACC TAA 3’

met gly ala leu stop


Stop codon mutation

A point mutation changing a stop codon into a codon

for an amino acid

5’ ATG GGA GCT CTA TTA ACC TAA 3’

met gly ala leu leu thr stop

5’ ATG GGA GCT CTA TTA ACC TTA 3’

met gly ala leu leu thr LEU
 Insertion or deletion mutations
 The genetic code is read in triplet nucleotides
during translation.

 Addition or subtraction of nucleotides not in

multiples of three lead to a change in the
reading frame used for translation. Amino
acids after that point are different, a
phenomenon called a frameshift.
Addition or subtraction of nucleotides in

multiples of three leads to addition or
subtraction of entire amino acids but not a
change in the reading frame.
 INSERTION

FRAMESHIFT MUTATION
 DELETION

FRAMESHIFT MUTATION
Molecular Effect of Mutation

Within coding region:

o Silent
o Missense
o Nonsense
o Frameshift

Within noncoding region:

o Splicing mutation
o Regulatory sequence mutation
Clinical Effect of Mutation

o Acceptable
o Partially acceptable
o Unacceptable (fatal/lethal)
 Mutations: Substitutions
Normal gene Substitution mutation
GGTCTCCTCACGCCA GGTCACCTCACGCCA
↓ ↓
CCAGAGGAGUGCGGU CCAGUGGAGUGCGGU
Codons
↓ ↓
Pro-Glu-Glu-Cys-Gly Pro-Arg-Glu-Cys-Gly
Amino acids
Substitutions will only affect a single codon
Their effects may not be serious unless they affect an amino acid that is
essential for the structure and function of the finished protein molecule
(e.g. sickle cell anaemia)
© 2010 Paul Billiet ODWS
 The genetic code is degenerate
A mutation to have no effect on the phenotype
Changes in the third base of a codon often have no
effect.

© 2010 Paul Billiet ODWS

 Mitochondrial code
U(Vertebrates)
C A G
UUU Phe UCU Ser UAU Tyr UGU Cys

U
UUC Phe UCC Ser UAC Tyr UGC Cys
UUA Leu UCA Ser UAA Stop UGA Trp
UUG Leu UCG Ser UAG Stop UGG Trp
CUU Leu CCU Pro CAU His CGU Arg

C
CUC Leu CCC Pro CAC His CGC Arg
CUA Leu CCA Pro CAA Gln CGA Arg
CUG Leu CCG Pro CAG Gln CGG Arg
AUU Ile ACU Thr AAU Asn AGU Ser

A
AUC Ile ACC Thr AAC Asn AGC Ser
AUA Met ACA Thr AAA Lys AGA Stop
AUG Met ACG Thr AAG Lys AGG Stop
GUU Val GCU Ala GAU Asp GGU Gly

G
GUC Val GCC Ala GAC Asp GGC Gly
GUA Val GCA Ala GAA Glu GGA Gly
GUG Val GCG Ala GAG Glu GGG Gly
Point mutations:
Single base mutations:

1. Missense mutation: leads to

an amino acid change

2. Silent mutation: does not

change the amino acid

3. Nonsense mutation: causes

premature stop-codon
 No change (Silent
Mutation)
Normal gene Substitution mutation
GGTCTCCTCACGCCA GGTCTTCTCACGCCA
↓ ↓
CCAGAGGAGUGCGGU CCAGAAGAGUGCGGU
Codons
↓ ↓
Pro-Glu-Glu-Cys-Gly Pro-Glu-Glu-Cys-Gly
Amino acids

© 2010 Paul Billiet ODWS

 Disaster
Normal gene Substitution mutation
GGTCTCCTCACGCCA GGTCTCCTCACTCCA
↓ ↓
CCAGAGGAGUGCGGU CCAGAAGAGUGAGGU
Codons
↓ ↓
Pro-Glu-Glu-Cys-Gly Pro-Glu-Glu-STOP
Amino acids

© 2010 Paul Billiet ODWS

 Mutations: Inversion
Inversion mutations, also, only affect a small part
of the gene
Normal gene Inversion mutation
GGTCTCCTCACGCCA GGTCCTCTCACGCCA
↓ ↓
CCAGAGGAGUGCGGU CCAGGAGAGUGCGGU
Codons
↓ ↓
Pro-Glu-Glu-Cys-Gly Pro-Gly-Glu-Cys-Gly
Amino acids

© 2010 Paul Billiet ODWS

 Mutations:
A frame shift mutation
Additions
Normal gene Addition mutation
GGTCTCCTCACGCCA GGTGCTCCTCACGCCA
↓ ↓
CCAGAGGAGUGCGGU CCACGAGGAGUGCGGU
Codons
↓ ↓
Pro-Glu-Glu-Cys-Gly Pro-Arg-Gly-Val-Arg
Amino acids

© 2010 Paul Billiet ODWS

 Mutations: Deletions
A frame shift mutation
Normal gene Deletion mutation
GGTCTCCTCACGCCA GGTC/CCTCACGCCA
↓ ↓
CCAGAGGAGUGCGGU CCAGGGAGUGCGGU
Codons
↓ ↓
Pro-Glu-Glu-Cys-Gly Pro-Gly-Ser-Ala-Val
Amino acids

© 2010 Paul Billiet ODWS

Crossing over:

• Chromatids change parts

between homologous chromatids
during the meiosis

• Causes redistribution of heridary

material between the homologous
chromosomes

 number of genes doesn’t

change
 new allele combinations
are formed
 CONSEQUENCES OF MUTATIONS
• CANCER
• INBORN ERRORS OF METABOLISM
• CHROMOSOMAL REARRENGEMENTS
(THE IMMUNE SYSTEM)
 CANCER
GENETIC CHANGES ASSOCIATED WITH CANCER:
1. ALTERED PROTEINS
2. ALTERED REGULATORY SEQUENCES
3. CHROMOSOMAL TRANSLOCATION
4. PROTO-ONCOGENE AMPLIFICATION
5. INSERTION OF VIRAL DNA INTO CHROMOSOMES
6. LOSS OR INACTIVATION OF ANTI-ONCOGENES
 Altered DNAAltered Protein
Cell
DNA Nucleus membrane

Chain of
amino
DNA acids
bases mRNA

Gene

Protein

Ribosome
 Hereditary Mutations
Egg Sperm

Mutation Occurs

Fertilized Egg Mutation

Reproductive Bone Pancreas Brain

Body Cells of Offspring
 Inherited diseases
• DNA mutations are significant in development of
diseases

• Inherited diseases are caused by mutations passed from

a parent to a offspring

• Monogenic diseases: disease is caused by one mutation

in one gene

• Multifactiorial diseases: disease is caused by interaction

of different mutations and environmental factors
 Mutations of haemoglobin
 Haemoglobin is a tetramer = 2  and 2 -chains
 The genes for these polypeptides are found on different
chromosomes
 The -chain gene is found on chromosome 11
 The -chain gene is found on chromosome 16
 The nucleotide sequences have been worked out
 Several inherited diseases occur on the -chain, which
contains 146 amino acids.

© 2010 Paul Billiet ODWS

 Mutation Codon Change to DNA Change in
sense strand Amino Acid
S (sickle cell 6 GAG to GTG Glu to Val
anaemia)
C (cooley’s 6 GAG to AAG Glu to Lys
syndrome)
GSan Jose 7 GAG to GGG Glu to Gly
E 26 GAG to AAG Glu to Lys
MSaskatoon 63 CAT to TAT His to Tyr
MMilwauki 67 GTG to GAG Val to Glu
OArabia 121 GAA to GTA Glu to Val

© 2010 Paul Billiet ODWS

 Hutchinson-Gilford’s Syndrome
Accelerated aging and show an early onset of aging: diabetes,
cataracts, osteoporosis, baldness and atherosclerosis
Mutation of a nuclear protein, Lamin A/C  involve in the
biology of the inner nuclear membrane
Impaired in DNA machinery
Suggest that efficient repair of DNA is essential for
prevention of Cancer and for normal aging.
 Changes in the DNA are important in normal aging
Hutchinson-Gilford’s Syndrome
 Two important terms...

Phenotype: The outlook of an organism

Genotype: The genetic information written in DNA

Phenotypes

Genotype Genotype

GCCAAGAATGGCTCCCACCT
ATGTTTCCACCTTCAGGTTCC
GGCTCTCAGACATTCCCCTG
ACTGGGCTGATTCCCCCCTCC
GTCCAACCCCCAGGCCATCA
CACTTTCAAGCTCGGCCCCTT
AGATGTCTCAGAGAGGCGG
TCAACTCAGAGAGGCGGCTA
CTAGACACCCAGAGACCTCA
GACACCCAGAGACCTCAAGT
AGTGACCATGTGGGAACGG
GACCATGTGGGAACGGGATG
GATGTTTCCAGTGACAGGCA
TTTCCAGTGACAGGCAG
DNA REPAIR
 DNA REPAIR
• DNA is a duoble-stranded  easier to be repaired
• RNA (HIV virus)  single strand, have a very high rate of
mutation
DNA REPAIR
MISMATCH REPAIR
PHOTOREACTIVATION
EXCISION REPAIR
 Two pathways of increasing complexity

Base
Excision
Nucleotide
repair
Excision
repair
 Base excision repair
Base excision repair pathway
Damaged base (BER).
(a) A DNA glycosylase recognizes
a damaged base and cleaves
between the base and deoxyribose
in the backbone.
(b) An AP endonuclease cleaves
the phosphodiester backbone
near the AP site.
(c) DNA polymerase I initiates
repair synthesis from the free 3’
OH at the nick, removing a
portion of the damaged strand
(with its 5’3’ exonuclease
activity) and replacing it with
undamaged DNA.
(d) The nick remaining after DNA
polymerase I has dissociated is
sealed by DNA ligase.

AP= apurinic or apyrimidinic

(a=without)
DNA repair defects cause disease
 HUMAN GENOME PROJECT
(HUGO)

• Mengidentifikasi semua gen dalam kromosom manusia

beserta fungsinya masing-masing
• Memetakan polimorfisme (variasi genetik) pada manusia
 HUMAN GENOME PROJECT
(HUGO)
• Genome manusia terdiri dari 3.164.700.000 pasang
basa
• < 2% mengkode protein
• Rata-rata ukuran gen 3000 bp
• Jumlah seluruh gen ± 30.000, yang baru diketahui
fungsinya ± 50%
• 1,4 juta SNP (single nucleotide polymorphisms)
Chromosome X
 SINGLE NUCLEOTIDE POLYMORPHISMS (SNP)

SPESIFIC REPEATED SEQUENCE (TANDEM REPEAT)