MUTATIONS AND DNA REPAIR MECHANISM

Mutations - alterations in gene sequence

- can be heritable if the mutation is on the germline (mutations
occurring on the DNA of cells that produce the egg and sperm)
- involves a change in the shape, structure or nucleotide sequence
of the DNA

Genetic disorders  a result of alterations of DNA sequences

DNA sequence  amino acid sequence  conformation  function

Mutations:
1. Spontaneous mutations
2. Induced mutations – caused by mutagens

Effects:
1. Lethal
2. Branching in the evolutionary tree or specie diversity

Type of Mutations according to Size:

1. Point Mutations - small or submicroscopic (only one or a small number of
nucleotides or bases are affected)
2. Gross Chromosomal Mutations -large-scale chromosome abnormality (seen
under the light microscope)

POINT MUTATIONS

A. SUBSTITUTION: single-base mutations, substitution of one or more

nucleotides by the same number of different nucleotides (in most cases,
only one)

2 types:
TRANSITION : purine to purine TRANSVERSION : purine to
pyrimidine
pyrimidine to pyrimidine pyrimidine to
purine

A T A T A T

G C G C G C

Consequences:

Mutation & DNA Repair Mechanism

 1. Silent mutation- no effect on the amino acid sequence of the protein
2. Missense mutation- results in the incorporation of a different amino acid in
the protein. The protein may be functional, partially functional or non-
functional
3. Nonsense mutation- results in the premature appearance of a stop codon
resulting in the shorter protein that is likely to be non-functional

DELETION: removal of one or more nucleotides in the DNA

*effect: frameshift mutation, a shift in the reading frame, which produces a
non-functional protein

B. INSERTION: addition of one or more nucleotides in the DNA

*if the number of inserted bases is not a multiple of 3, it will cause frameshift,
resulting in serious consequences
*some diseases caused by insertion: Huntington disease, Kennedy disease,
Myotonic dystrophy, etc.
*deletion and insertion are usually caused by Replication Slippage

C. EXON SKIPPING: results from mutation at the splice site

*Splicing of an intron requires an essential signal: for example, "GT........AG". If
the splice acceptor site AG is mutated, the splicing machinery will look for the
next acceptor site. As a result, the exon between two introns is also removed.

Mutation & DNA Repair Mechanism

 SPONTANEOUS MUTATIONS

How do spontaneous mutations occur?

 As part of normal DNA replication – DNA replication is not an error free

process (proof-reading ability of DNA polymerase)

 Insertion/Deletion of one or more nucleotides as a result of a slippage error

during DNA replication

 TAUTOMERIC SHIFTS
o rare and reversible redistributions of electrons and protons in the
structures of the bases in the DNA. This alters their base pairing
properties.
o can cause spontaneous mutation when the bases convert to their rare
tautomeric forms just as the bases are being copied during replication or
transcription
o the tautomeric forms of the bases are very unstable hence short-lived
o the probability of changes due to spontaneous mutations is very low.
Since changes in the genetic make-up of a species depend on
spontaneous mutations, molecular evolution is very slow
o tautomers: keto and enol, amino and imino

Mutation & DNA Repair Mechanism

 INDUCED MUTATIONS

Induced mutations occur more frequently than spontaneous mutations.

A. Mutations due to PHYSICAL AGENTS in the form of high energy radiation

 UV light leading
to the formation of THYMINE
DIMERS (covalent linkage of
2 adjacent thymines on the
same strand of DNA) which
can block DNA replication or
interfere with base pairing.

 Ionizing radiation such as X-rays leading to single and

double-stranded DNA breaks or to the formation of OH-radicals from
water

B. Mutations due to CHEMICAL / ENVIRONMENTAL SUBSTANCES

 BASE ANALOGUES
- have similar structures to the DNA bases
- more prone to tautomeric shifts

Example: 5-Bromouracil
O O

Br C H3C C
5-Bromouracil is a structural
C NH C NH analogue of thymine. It
HC C
undergoes tautomeric shift to
HC C
NH O NH O base pair with guanine
5-bromouracil thymine instead of adenine.
 ALKYLATING AGENTS
-adds a methyl or ethyl group to a base
-the largest class of “potential” mutagens present in man’s environment

example: N-nitrosoamines (found in cigarette smoke)

When N-nitrosoamine is in the liver, it is metabolized by liver enzymes to

form alkylating agents which can attack Guanine. There are 2 reactive
sites in G, the N7 and O6 positions. If alkylation occurs at the N7, the
reaction is reversible due to the presence of “suicide” enzymes called
demethylase. If alkylation occurs at the O6 position, an apurinic site in the
DNA is formed. During replication, an apurinic site may be ignored

Mutation & DNA Repair Mechanism

 resulting in a deletion in the daughter strand or a base is selected at
random and placed in the daughter strand.

 DEAMINATING AGENTS
-removes amino groups from a base

Example: Sodium nitrite NaNO2 (used as a preservative,

color enhancer and color fixative in bacon, smoked fish,
tocino, etc.)

When ingested, sodium nitrite is converted to nitrous

acid (HNO2) in acidic conditions. Nitrous acid removes
groups from adenine, guanine and cytosine. For
example, deamination of adenine results in the
formation of hypoxanthine, a structural analogue of guanine hence may
base pair with cytosine resulting in transition.

Example: Cytosine to Uracil

 INTERCALATING AGENTS
-contains a cyclic system that can interact with the bases of DNA
-do not chemically modify DNA but physically binds to it by becoming
inserted between adjacent base pairs because of their flat ring structures.
This affects the opening of the DNA during replication or transcription.

Example: Benzopyrene (found in automotive exhaust & cigarette smoke),

Benzene (an organic solvent), Aflatoxin (a metabolic product of molds in
peanuts, oils & grains)

 VIRAL AGENTS

Mutation & DNA Repair Mechanism

 -some viruses contain oncogenes (cancer-causing genes) which can be
activated once they insert their DNA in the host’s genome (the process is
called lysogeny).
-the virus that infects a cell may either undergo lytic or lysogenic cycle
-when viruses insert their DNAs into the host genome, the sequence of
the bases of the host DNA may be altered or certain destructive genes
can be activated. (Figure on last page)

REPAIR MECHANISMS
A. ENZYMES
Examples:
*Photoreactivating enzyme which recognizes thymine dimers and
monomerizes it upon absorption of visible light.
*DNA glycosylases recognize altered bases and catalyze its hydrolytic
removal from deoxyribose.

B. FREE-RADICAL SCAVENGERS / ANTI-OXIDANTS

Examples:
*Glutathione and Metallothionein
*Enzymes such as superoxide dismutase, catalase, and peroxidases
*Vitamins such as Vitamins C, A and E (conjugated double bonds in their
structures react with free radicals)

C. DARK REPAIR MECHANISM

• Excision repair
-example:
thymine dimers in one strand only
Post replication repair
-example:
thymine dimers in 2 strands

LYTIC CYCLE ( ) / LYSOGENIC CYCLE ( )

Mutation & DNA Repair Mechanism

 Binary Fission

Mutation & DNA Repair Mechanism