MUTATION

INTRODUCTION
• MUTATION- Any sudden change
occurring in hereditary material is
called as mutation.
• They may be harmful, beneficial or neutral.
• DNA is highly stable molecule that replicates
with amazing accuracy , some errors of
replication do occurs.
 It can also be defined as sudden and
spontaneous changes in the cell. Mutations are
caused by radiation, viruses, transposons and
mutagenic chemicals, as well as errors that
occur during meiosis or DNA replication. They
can also be induced by the organism itself, by
cellular processes such as hyper mutation.
 Due to the damaging effects that mutations can
have on genes, organisms have mechanisms
such as DNA repair to remove mutations.
 Viruses that use RNA as their genetic material
have rapid mutation rates, which can be an
advantage since these viruses will evolve
constantly and rapidly, and thus evade the
defensive responses of e.g. the human
immune system.
 HISTORY
• 1901: Hugo de Vries first used
the term mutation to describe
the sudden heritable phenotypic
changes in evening primrose
Oenothera lamarckiana.
• 1904: T.H. Morgan reported
white eyed drosophila in the
population of red eyed flies.
• 1928: H.J. Muller first used x-
rays to induce mutation in fruit Thomas
fly. Hunt Morgan

Masatoshi
Definition…..
 Can occur in somatic cells or within germ
cells
 Germ cell mutations –
 heritable
 basis for the transmission of genetic diversity
and evolution as well as genetic diseases
 Somatic cell mutation –
 nottransmitted to the next generation
 may lead to altered cellular functions or tumors
Other terms
 Mutagen – a physical or chemical agent
that causes mutation to occur (or increases
the frequency of their occurrence)
 Mutant – an organism or a gene that is
different from the normal or wild type
 Mutagenesis – process of producing a
mutation
 TYPES OF MUTATIONS
 The sequence of a gene can be altered in a number of ways.
Gene mutations have varying effects on health depending on
where they occur and whether they alter the function of
essential proteins. Mutations in the structure of genes can be
classified as:
Types of gene mutation:
1. Point mutation
2. Insertion mutation
3. Deletion mutation
4. Frame shift mutation
 Point mutations, often caused by chemicals or malfunction of
DNA replication, exchange a single nucleotide for another.
 Point Mutations are of two types:
1.Transitions
2.Transversions.
 Most common is the transition that
exchanges a purine for a purine (A ↔ G) or
a pyrimidine for a pyrimidine, (C ↔ T). A
transition can be caused by nitrous acid,
base mis-pairing, or mutagenic base
analogs such as
5-bromo-2-deoxyuridine (BrdU).
 Less common is a transversion, which
exchanges a purine for a pyrimidine or a
pyrimidine for a purine (C/T ↔ A/G). An
example of a transversion is adenine(A)
being converted into a cytosine (C).
TRANSITION
MUTATION

A transition is the
replacement of a base by
the other base of the
same chemical.

Griffiths et al.,
2002
TRANSVERSION MUTATION

A transversion is the
opposite the
replacement of a base
of one chemical
category by a base of
the other.

Griffiths et al.,
 Point mutations that occur within the protein
coding region of a gene may be classified
into three kinds, depending upon what the
erroneous codon codes for:

 Silent mutations

 Missense mutations

 Nonsense mutations
 Silent mutations: which code for the same amino acid.
 Missense mutations: This type of mutation is a change in one
DNA base pair that results in the substitution of one amino acid for another
in the protein made by a gene.
 Nonsense mutations: A nonsense mutation is also a
change in one DNA base pair. Instead of substituting one amino acid for
another, however, the altered DNA sequence prematurely signals the
cell to stop building a protein. This type of mutation results in a
shortened protein that may function improperly or not at all.
 Insertions add one or more extra
nucleotides into the DNA. They
are usually caused by
transposable elements, or
errors during replication of
repeating elements (e.g. AT
repeats). Insertions
in the coding region of a gene may
alter splicing of the mRNA (splice site mutation), or cause a
shift in the reading frame (frameshift), both of which can
significantly alter the gene product. Insertions can be
reverted by excision of the transposable element.
 Deletions remove one or
more nucleotides from the DNA.
Like insertions, these mutations
can alter the
reading frame of the gene. They
are generally irreversible: though
exactly the same sequence might
theoretically be restored by an insertion,
transposable elements able to revert a
very short deletion (say 1–2 bases) in any location are
either highly unlikely to exist or do not exist at all. Note
that a deletion is not the exact opposite of an insertion:
the former is quite random while the latter consists of a
specific sequence inserting at locations that are not
entirely random or even quite narrowly defined.
 Frame shift mutation: A frameshift mutation is a mutation
caused by insertion or deletion of a number of nucleotides that is not evenly
divisible by three from a DNA sequence. Due to the triplet nature of gene
expression by codons, the insertion or deletion can disrupt the reading frame,
or the grouping of the codons, resulting in a completely
different translation from the original.The earlier in the sequence the deletion
or insertion occurs, the more altered the protein produced is.
 Mutations in chromosomal structure, including
 Changing the structure of chromosome.
 Loss or gain part of a chromosome.

 Chromosomal Mutation five types exist:

1.Deletion
2.Inversion
3.Transloaction
4.Duplication
5.Non Disjunction
 Deletions of large chromosomal regions,
leading to loss of the genes within those
regions
 INVERSION
 Chromosome segment breaks off
 Segment flips around backwards
 Segment reattaches
 Translocations: Interchange of genetic parts from
non homologous chromosomes.

 Involves two chromosomes that are not homologous.

 Part of one chromosome is transferred to another
chromosomes
 Duplications: leading to multiple copies of all
chromosomal regions, increasing the dosage of the genes
located within them.
 Nondisjunction
 Failure of chromosomes to separate during
meiosis
 Causes gamete to have too many or too
few chromosomes
 Disorders:
 Down Syndrome -
 Turner syndrome-
 Klinefelter’s syndrome-
Type of Mutagenic Agents
 Base analogue
 Nitrous acid
 Hydroxylamine
 Alkylating agents
 Intercalating agents

 UV radiations
Base analogue
 Compounds that can substitute for purines
and pyrimidines during nucleic acid
biosynthesis
 Example: 5-bromouracil – derivative of
uracil; behaves as a thymine analog
 Presence of bromine in place of methyl
group increases the probability that a
tautomeric shift will occur
Base analogue…
 If 5-BU incorporated into DNA in place of
thymine and a tautomeric shift to enol
occurs – 5-BU base pairs with guanine
 After one round of replication – an A=T to
G=C transition results
 Presence of BU increases the sensitivity
of the molecule to UV light, a known
mutagen
Other examples of base
analogues
 In wheat about a quartet of the cytosine fraction
exists as the 5-methyl cytosine.
 5-methyl cytosine also occur naturally in DNA of
grasses and many mammals.
 In T-even phages of E. coli cytosine is completely
replaced by 5-hydroxylmethyl cytosine and 5-
glucosyl hydroxymethyl cytosine.
 Other naturally occurring base analogues include 5-
hydroxylmethyl uracil which occurs in certain viruses
and 6-methyl purine, an adenine analogue, which
occurs in some bacteria.
Nitrous acid
 Formed by digestion of nitrites
(preservatives) in foods
 Very potent mutagen that acts directly on
either replicating or non replicating DNA
by oxidation or deamination of the bases
that contain amino groups (adenine,
guanine and cytosine)
 Conversion of the amino groups to keto
groups changes the hydrogen bonding
potential of the bases
Nitrous acid…
 Adenine is deaminated to hypoxanthine,
which then pairs preferentially with cytosine
in the place of thymine
 Cytosine is deaminated to uracil, which now
pairs with adenine in the place of guanine
 Deamination of guanine has zero effect, as
deaminated guanine also pairs with cytosine.
Nitrous acid…
 Deamination of adenine leads to AT GC
transition and the deamination of cytosine
results in CG AT transitions, nitrous acid
induces transitions in both directions
 Nitrous acid also causes interstrand cross-
linking of DNA
 DNA strands fail to separate and there is no
DNA duplication, which is lethal or
deleterious
Hydroxylamine
 Hydroxylamine (NH2OH) reacts with pyrimidine
bases
 Its effect is strong on cytosine while on uracil the
effect is less
 Hydroxylamine breaks and removes pyrimidine ring
of uracil thus producing phosphoribosyl urea and 5'­
isoxasolone with cytosine
 Hydroxylamine finally produces hydroxyl amino (-
NOH) derivative, which might be responsible for
base pair change (GC-AT pair)
 Mutations induced by hydroxylamine cannot be
reversed with hydroxylamine
Alkylating agents
 Alkylating agents carry one, two or more
alkyl groups in reactive form, which are
capable of being transferred to other
molecules where electron density is high
 One major mechanism ,of mutagenesis by
alkylating agents involves the transfer of
methyl or ethyl group to the bases such
that their base-pairing potentials are
altered and transitions result
Alkylating agents…
 Ethylmethane sulfonate (EMS, a mustard gas) is
an alkylating agent that can donate an alkyl group
(C2H5) to a keto group in guanine
 Resulting 6-ethylguanine acts as an analog
of adenine and pairs with thymine, leading
to transition mutations.
 Result is transition of GC to AT.
 Alkylating agents work by three different
mechanisms all of which achieve the same end
result - disruption of DNA function and cell death.
Alkylating agents…
 In the first mechanism an alkylating agent
(represented in the figure below as a pink star)
attaches alkyl groups (small carbon compounds-
depicted as pink triangles) to DNA bases
 This alteration results in the DNA being
fragmented by repair enzymes in their attempts
to replace the alkylated bases (frame 3 of the
diagram below)
 Alkylated bases prevent DNA synthesis and
RNA transcription from the affected DNA
Alkylating agents…
 A second mechanism by which alkylating agents
cause DNA damage is the formation of cross-
bridges, bonds between atoms in the DNA (pink
linkages below)
 In this process, two bases are linked together by an
alkylating agent that has two DNA binding sites
 Bridges can be formed within a single molecule of
DNA (as shown below) or a cross-bridge may
connect two different DNA molecules. Cross-linking
prevents DNA from being separated for synthesis or
transcription.
Alkylating agents…
 The third mechanism of action of alkylating
agents is the induction of mispairing of the
nucleotides leading to mutations.
 In a normal DNA double helix, A always pairs
with (is across from) T and G always pairs with
C. As the figure below shows, alkylated G
bases may erroneously pair with Ts.
 If this altered pairing is not corrected it may
lead to a permanent mutation.
Alkylating agents…
 Alkylating agents may readily react with
PO4 group and may break the sugar
phosphate backbone of DNA.
 This may induce large alterations.
Difunctional alkylating agents cause cross
linking of DNA, induce chromosome breaks
and chromosomal aberrations.
 Alkylating agents as a class exhibit less
specificity in their mutagenic effect than
base analogs or dyes.
Alkylating agents…

 They induce all types of mutations,

(transitions, transversions, frameshifts and
even chromosome aberrations) with
various relative frequencies depending on
the specific alkylating agent.
UV radiations
 Purines and pyrimidines absorb UV radiation
most intensely at a wavelength of about 260 nm.
 One of the major effect of UV radiation – creation
of pyrimidine dimers – chemical species
consisting of two identical pyrimidines –
particularly one consisting of two thymine
residues
 Dimers distort the DNA conformation and inhibit
normal replication
UV radiations…
 As a result, errors can be introduced in the
base sequence of DNA during replication.
 When UV-induced dimerization is
extensive, it is responsible for the killing
effects of UV radiation on cells.
Mutator Genes
Discovered recently
Repair the damaged DNA
In E. coli there are genes that, when present in a
mutant state, cause mutations to appear very
frequently in other genes throughout the genetic
map – mutator genes
Only when the product of the mutator gene is itself
defective will there be widespread production of
mutations
Any gene that produces an increase in
spontaneous mutation rates
 Map
Locus Position Specificity Strength Defect (if known)
(min)
Prevents incorporation of
A:T->C:G
mutT 2 Moderate A:8-oxodG mispairs by
transversions
hydrolyzing 8-oxo-dGTP
All base
altered e subunit of DNA
mutD 5 substitutions very strong
polymerase III
frameshift
Lacks glycosylase that
G:C->T:A
mutY 64 Moderate corrects G:A and 8-
trasnversion
oxodG:A mispairs
G:C->T:A Fapy glycosylase (8-
mutM 82 Moderate
trasnversion oxodG glycosylase)
A:T-> T:A, G:C-
>T:A,and A:T-
mutA 95 Moderate Not Known
>C:G
transversions
A:T-> T:A, G:C-
>T:A,and A:T- Weak/
mutC 42 Not Known
>C:G Moderate
transversions
Mutator Gene…..
 Their action may be limited to certain alleles or
to certain mutational pathways
 Four types that yield strong phenotypic effects:
 A mutant DNA polymerase that reduces or
eliminates the 3’-5’ exonuclease activity of the
editing function
 A mutant methylating enzyme (the dam enzyme)
responsible for methylation of the sequences that
the mismatch repair system uses to discriminate
parental strands from daughter strands
Mutator Genes…..
A mutant enzyme that cannot carry out the
excision step of mismatch repair
 Mutations in the regulatory circuits that
maintain the error prone SOS repair system is
an “off” state
Mutator Genes…..
 Hereditary nonpolyposis colon cancer (HNPCC) is
caused by mutations in one of the five mutator genes,
known so far
 MSH2 - locus frequently mutated in hereditary
nonpolyposis colon cancer (HNPCC)
 When cloned, it was discovered to be a human homolog
of the E. coli mismatch repair gene mutS, consistent
with the characteristic alterations in microsatellite
sequences (RER+ phenotype) found in HNPCC.
 Also associated with some endometrial cancers
Reversion
 It is the reverse process, in which wild-type is
regained
 Also called as back mutation or reverse
mutation
 May occur spontaneously or may be induced
 Mechanisms:
 Back mutation occurs at the same site as the
original mutation and restores the wild-type base
sequences – true reversion
Reversion…
 Wild-type phenotype is restored as a result of
a second mutation occurring at a different site
– pseudoreversion, second-site or suppressor
mutation
 May be situated within the same gene
(intragenic suppression) or within a different
gene (intergenic suppression)
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