MUTATION

Dr.H.B.Mahesha
Associate Professor and Head
Department of Sericulture
Yuvaraja’s College
University of Mysore, India.
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 Significance of Mutations
According to one hypothesis mutation is responsible for the separation of
existing human beings from our ancestor primates
Future

Present day human

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Mutation is an abrupt qualitative or quantitative change in the genetic material/
Any change in the nucleotide sequence of DNA, which gives rise to a mutant genotype

The organism which undergoes mutation is known as mutant

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 History
The earliest record of
point mutation dates back
to 1791, when Seth Wright
noticed a lamb with
exceptionally short legs
in his flock of sheep.

Mutation - Coined by Hugo de Vris in1900 to explain the heritable changes

in evening primrose Oenothera lamrckiana

Mutagenesis - Process of producing mutations

Mutagen - A physical/chemical/biological agent that causes mutations

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The first scientific study of mutation started in 1910, when
Morgan started his work on fruit fly Drosophila
melanogaster after he observed white eyed male individuals
among red eyed male individuals.

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Classification
Based on the origin: Natural / Artificial (induced)
Based on the type of cells: Somatic/Gametic
Based on type of chromosomes: Autosomal / Allosomal
Based on direction: Forward/Reverse
Based on size: Point/Gross
Silent Mutations:- has no detectable effect of phenotype
Leaky mutations:- Causes amino acid substitution, eventually it
reduces the activity of an enzyme.
Nonsense Mutations or Chain termination Mutation:- Causes
premature termination of polypeptide chain.
etc.,

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Natural Mutation
Spontaneous mutations occur suddenly in the nature and
their origin is unknown. They are also called as background
mutations and have been reported in many organisms such
as Oenothera, Maize, Drosophila, Mice, Man etc.,

For example TAUTOMERISM

Frequency of total mutations - 10-7 to 10-12/organism

(i.e., 0.0000001- 0.000000000001)
Frequency of detectable mutations - 1 in 106 (i.e., 0.000001)

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 Tautomerism:-The ability of a molecule to exist in more
than one chemical form

Normal DNA bases and their pairing patter

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 Regular and Tautomeric forms of DNA Bases

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 All the four common bases of DNA (adenine, guanine,
cytosine and thymine) have unusual tautomeric forms,
which are however rare. A tautomeric shift is believed
to occur when an amino (NH2) form of adenine is
changed to an imino (NH) form.

Base change from AT to GC

MUTATION

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 Similarly a tautomeric shift may occur in thymine
changing from keto (C=O) form to rare enol (COH)
form. Natural base pairing in DNA is A=T and G C.
The taustomeric forms are however capable of
unusual (forbidden) base pairing like T G, G T,
C=A and A=C.

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 Induced Mutation
Mutation can be induced artificially in the living
organisms by exposing them to abnormal
environment such as radiations, certain
physical conditions (i.e. temperature) and
chemicals.

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 Mutagens
The substances or agents which induce mutations
are called mutagens or mutagenic agents. They
may be physical, chemical or biological.

1.Physical –Radiations: i. Ionizing-X-rays, gamma, Alpha,

Beeta, protons, nutrons etc.,

ii. Non Ionizing -U V

Temperature-

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 2.Chemical –
Base Analogues: 2,aminopurine, 5-bromourasil etc.,
Base Modifying agents: Nitrous acid, Hydroxylamine etc.,
Distortion producing agents: Proflavin, acridine orange etc.,

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 3.Biological Mutagens: They may be viral or
bacterial.
H. Pylori- implicated in stomach cancer
Hepatitis B virus- implicated in liver cancer
H. papiloma virus-implicated in cervical cancer
Human T-cell lymphocytic virus implicated in
lymphoma

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 Point Mutation / Gene Mutation
A mutation that changes only one small area or one
nucleotide in a gene
Earliest record of point mutation dates back to
1791 by Seth Wright.
Since then mutations have been reported in E.
coli, Neurospora, Pea, Maize, Rodents, Fowls,
Man, etc.,

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 Frame Shift Mutation
A mutation that inserts or deletes a single base
(normally single rarely more) will change the reading frame

for the entire subsequent sequence.

A change of reading frame is called Frame shift
mutation.

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 Central Dogma of Molecular Biology
Shows the connection between DNA, RNA & Protein

As shown in this
picture,
If any change
occurs in DNA
Molecule; that
alters the
reading frame
during
Transcription
and translation.
Hence, it is
called as Frame
shift mutation

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 Frame Shift Mutation may be
i. Deletion: Removal of one or few bases from a nucleotide
chain is called a deletion.
ii. Insertion: This is due to the addition of one or more (few)
nucleotides to the DNA.

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 Substitution:- A nitrogenous base of a
codon is replaced by another base is
called substitution mutation. They maybe
1. Transition
2. Transversion

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 1. Transition: When a purine base of a triplet codon is
substituted by another purine base or a pyrimidine
base is substituted by another pyrimidine

Base change from AT to GC

MUTATION
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 Transversion: The substitution mutation when
involves the substitution or replacement of a purine
with a pyrimidine or vice versa, then such type of
substitution mutation is called transversion
mutation.

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Chemical Mutagenesis
Test for mutagenic effects of chemical agents are
almost as old as modern genetics. In 1934, Morgan
tried to produce mutations in Drosophila by
treating with alcohol and ether, but without
success. After a number of attempts by many
workers the search for chemical agents met with
only during Second World War.
Types:
Copy Errors by Base Analogues
Direct Effect on DNA
Agents Producing Distorsions in DNA

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Copy Errors by Base Analogues: These base analogues
have the molecular structure similar to that of nucleic
acid bases, which are incorporated in to DNA without
destroying its capacity for replication. However,
because the analogue differs from the normal base in
the distribution of hydrogen atoms, it has greater
tendency for improper pairing and causes mutation.

Eg.,

Thymine 5 Bromouracil

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 If 5-bu in enol state is incorporated in to DNA, in enol state it pairs with guanine and in the
next generation in keto state it pairs with adenine. Thus the G C pair is replaced by A=T
pair. This process occurs at replication and hence called replication errors.

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 Direct Effect on DNA
Agents modifying purines or pyrimidine include Nitrous acid (HNO2),
Hydroxylamine (NH2OH), Alkylating agents

Nitrous acid (HNO2):

it is a very powerful
mutagen becauses it
acts directly on the
nucleic acid,
replacing amino
groups (NH2) by
hydroxyl groups
Guanine Uracil =Adenine (OH).

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Hydroxylamine NH2OH: It reacts specifically with cytosine and
converts it to a modified base that pairs only with adenine so
that a G C pair ultimately becomes an A = T pair
Cytocin --------->Modified
(NH OH) 2
Base pairs with Adenine

Alkylating Agents: Like ethyl methane sulfonate (EMS), ethyl

ethane sulfonate (EES) have been used extensively in genetic
engineering research. These alkylating agents impair the normal
hydrogen bonding of the bases causing mispairing of G with T, leading
to transition of A=T > G C and G C to A=T. also induces recessive
lethal mutations, specific locus mutation, translocations, dominant
lethal and partial and complete chromosomal loss in Drosophila
melanogaster. EMS causes lethal mutations, deletion, translocation,
dominant lethal in silkworm Bombyx mori.

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 Agents Producing Distortions in DNA
Certain fluorescent acridine dyes like proflavin
and acridine orange causes mutations by insertion
or deletion of bases. The acridines are planer (flat)
molecules like purine bases and can be
intercalated between the bases of the DNA helics.
This distorts the structure of DNA. Result in
deletion or insertion of bases during replication.

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 Intercalation resulting in addition of the base

Acridine

x
Acridine
X-Unknown Base

Intercalation of the acridine molecule a base (x’) is inserted at random

opposite the acridine molecule
in the new chain.

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RADIATION MUTAGENESIS
Ionizing radiations:- High-energy radiation
capable of producing ionization in substances
through which it passes.
Ionization:- Any process that leads to the
dissociation of a neutral atom, molecule or
other species into ions; the state of being
ionized.
Eg., X-rays, α, β, gamma rays, fast moving
particles etc.,
Non Ionizing radiations:- Uv, Radio waves,
Visible Light
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 Ionizing

Besides ionizing radiations causes chromosome type as well as chromatid type

aberrations namely terminal deletion, interstitial deletion, translocation,
inversion etc., this can be seen under a compound microscope.

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 Effect of Non Ionizing radiation on DNA molecule

UV light causing nitrogen bases to become highly

reactive free radicals. The resulting unstability
causes conversion of one base to another (a purine
to another purine or a pyrimidine to another
pyrimidine).

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 The primary mutagenic effect of UV light is the
production of dimmers. Irradiation of a bacterial culture
and subsequent extraction of DNA yields three possible
types of pyrimidine dimmers in DNA. That is T=T at 50%;
T=C at 40% C=C at 10%

Thimine Dimers

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 Thymine Dimer formation and Repair mechanism

Thymine cytosine Dimer formation and Repair mechanism

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 Acknowledgements
to
1. Internet
2. Molecular biology by David Freifelder
3. Cell Biology by C B Powar

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