Shikha Yashveer1, Jayanti Tokas2, Shalini Jain3 and Hariom Yadav4

1
Department of Molecular Biology and Biotechnology, 2Department of Biochemistry, CCS HAU,
Hisar, Haryana, India
3
Department of Biochemistry, PGIMER, Chandigarh, India
4
National Agri-Food Biotechnology, Mohali, Punjab, India
Email: yadavhariom@gmail.com
  Any sudden change occurring in
hereditary material is called as
mutation

They may be harmful, beneficial or neutral

 In multicellular organism, two broad
categories of mutations: Somatic
mutations & germ line mutations
 Somatic mutations

Arise in the somatic cells

Passed on to other cells through the process
of mitosis
Effect of these mutations depends on the
type of the cell in which they occur & the
developmental stage of the organism
If occurs early in development, larger the
clone of the mutated cells
Germ line mutations

 They occur in the cells that produce

gametes
 Passed on to future generations
 In multicellular organisms, the term
mutation is generally used for germ
line mutations
Some Facts

 Term mutation was given by Devries

in 1901 while studying evening
primerose Oenothera lamarckiana
 Most of these were chromosomal
variations
 Some were point variations
 Originally the term mutation was
given to both chromosomal as well as
point mutations
Cont.

 Recently chromosomal mutations

are studied separately
 The term mutation is now given only
to point mutations
Definition

 DNA is a highly stable molecule that

replicates with amazing accuracy
 Some errors of replication do occur
 A mutation is defined as an inherited
change in genetic information
Types of gene mutation
Number of ways to classify gene
mutations:
 On the basis of the molecular nature of the
defect
 On the nature of the phenotypic effect--
amino acid sequence of the protein is
altered or not
 On the basis of the causative agent of the
mutation
 Base substitution
 Insertions & deletions
Base substitution:

 Simplest type of gene mutation

 Involves the alteration of a single
nucleotide in the DNA
 A base substitution usually leads to base pair
substitution
GGG AGT GTA GAT
CGT
CCC TCA CAT CTA GCA
GGG AGT GCA GAT
A base substitution
CGT
CCC TCA CAT CTA GCA
First cycle of DNA replication

GGG AGT GCA GAT

CCC TCA CAT CTA GCA
CGT
GGG AGT GTA GAT
CCC TCA CGT CTA GCA
CGT
Base substitution is of two types:

Transition:
Purine is replaced with a purine

Pyrimidine is replaced with a pyrimidine

Insertions & deletions:

 2nd major class of gene mutation

 Addition or the removal, respectively, of
one or more nucleotide pair
 Usually changes the reading frame, altering
all amino acids encoded by codons
following the mutation
 Also called as frame shift mutations
 cont.

Additions or deletions in the multiples of three

nucleotides will lead to addition or deletion of one or
more amino acids

These mutations are called in-frame insertions and

deletions, respectively.
Mutations on the basis of the Phenotypic effects of
mutations:

Most common phenotype in natural populations of the

organism is called as wild type phenotype

 The effect of mutation is considered with reference

to wild type phenotype
 Forward mutation:
 a mutation that alters the wild type
phenotype

Reverse mutation (reversion):

 a mutation that changes a mutant
phenotype back in to the wild type
Missense mutation: a base is substituted that alters a
codon in the mRNA resulting in a different amino acid in
the protein product

TCA TTA
AGT AAT

UCA UUA

Ser Leu
Nonsense mutation: changes a sense codon into a
nonsense codon. Nonsense mutation early in the mRNA
sequence produces a greatly shortened & usually
nonfunctional protein

TCA TGA
AGT ACT

UCA UGA Stop codon

Ser
Silent mutation: alters a codon but due to degeneracy of
the codon, same amino acid is specified

TCA TCG
AGT AGC

UCA UCG

Ser Ser
 Neutral mutation: mutation that alters the amino acid
sequence of the protein but does not change its function as
replaced amino acid is chemically similar or the affected aa
has little influence on protein function.

CTT ATT
GAA TAA

CUU AUU

Leu Ile
 Loss of function mutations:

Complete or partial loss of the normal function

Structure of protein is so altered that it no longer
works correctly
 Mutation can occur in regulatory region that
affects transcription , translation or spilicing of
the protein
 Frequently recessive
 Gain of function mutations:

Produces an entirely new trait

Causes a trait to appear in inappropriate tissues or at
inappropriate times in development
Frequently dominant
 Conditional mutations:
Expressed only under certain conditions

Lethal mutations:
 Cause the death of the organism
 Suppressor mutation:
Suppresses the effect of other mutation
 Occurs at a site different from the site of original
mutation
 Organism with a suppressor mutation is a double
mutant but exhibits the phenotype of un mutated
wild type
 Different from reverse mutation in which mutated
site is reverted back into the wild type sequence
 On the basis of Causative agent of mutation:

Spontaneous:
 Mutations that result from natural changes in
DNA

Induced:
 Results from changes caused By
environmental chemicals & radiations
 Any environmental agent that increases
the rate of mutation above the
spontaneous is called a mutagen such as
chemicals & radiations
 Chemical Mutagens:
 First discovery of a chemical mutagen was made by
Charlotte Auerbach

Base Analogs:
Chemicals with structures similar to that of any of the four
standard bases of DNA
DNA polymerases cannot distinguish these analogs
They may be incorporated into newly synthesized DNA
molecules
 5-bromouracil
an analog of thymine

O O

4 4
N 3 5 Br N 3 5 CH₃
5BU T
2 2
6 6
O 1 O 1
N N
 OH
O

4 4
N 3 5 Br N 3 5 Br
5BU 5BU
2 2
6 6
O 1 O 1
N N

Keto Enol
pairs with A mispair with G
 T TRANISITION
A T C
5dBU A G
5dBU
A

5dBU
G

C
G
 3’ 5’ 3’ 5’
GAC
GAC CTG
3’ 5’ 5’ 3’
GAC
3’ 5’ CBG
GAC CBG 5’ 3’
5’ 3’ CBG
5’ 3’
3’ 5’ Incorporated error GGC
GAC Strand 3’ 5’
CTG seperation 3’ 5’
5’ 3’ 5’ 3’
CBG GGC
3’ 5’
GAC 3’ 5’ 3’ 5’
CTG CTG GAC GGC
5’ 3’ 5’ 3’ CBG CCG
replication 5’ 3’ 5’ 3’
 G TRANISITION
C
G A
5dBU
C T
G
5dBU

5dBU
A

A
T
2-amino purine (P)

 Base analog of adenine

 Normally pairs with thymine
 May mispair with cytosine
 Causes a transition mutation
 T.A C.G
Incorporated error
3’ 5’ 3’ 5’
3’ 5’ 3’ 5’ GTC GTC
GTC GTC CAG
3’ 5’
GTC CPG 5’ 3’
Strand 5’ 3’ 5’ 3’
CAG CPG CPG
separation 3’ 5’ 5’ 3’
5’ 3’ GTC GCC
CAG 3’ 5’
5’ 3’ CAG
5’ 3’ 5’ 3’ 3’ 5’
CPG GCC
replication
3’ 5’ 3’ 5’
GTC GCC
CAG CGG
5’ 3’ 5’ 3’
T TRANISITION
A T C
2AP
A G
T
2AP

C
2AP

C
G
C TRANISITION
G C T
2AP
G A
C
2AP

T
2AP

T
A
Both base analogs produce transition
mutations
Mutations by base analogs can be
reversed by treatment with the same
analog or different analog
Alkylating agents:
Chemicals that donate alkyl groups e.g.
ehylmethanesulfonate(EMS)
It adds an ethyl group to guanine and produces 6-
ethylguanine, which pairs with thymine and leads to
CG:TA transitions
Also adds an ethyl group to thymine to produce 4-
ethylthymine, which then pairs with guanine, leading to a
TA:CG transition
Mutations produced by EMS can be reversed by
additional treatment with EMS.
Mustard gas is another alkylating agent.
C T
G A
EMS EMS

T 4ET
6EG G

T C
A G
 Nitrous acid: causes deamination
Cytosine Uracil

NH2 o

4 4
N 3 5 N 3 5

2 HNo2 2
6 6
O 1 O 1
N N
H H
CYTOSINE URACIL
5’ 3’
C HNO2
G 5’ 3’
3’ 5’ 5’ 3’ U
5’ 3’ U A
U 3’ 5’

3’
G 5’ 5’ 3’ 3’ 5’
G U A
3’ 5’

5’ 3’ 5’ U 3’ 3’ 5’
C
G A TA
3’ 5’ 3’ 5’ 5’ 3’

C.G TA
Adenine changes into Hypoxanthin which then pairs with Cytosine

5’ 3’
A
HNO2 5’ 3’
T 5’
3’ 5’ 3’ H
5’ 3’ H C
H 3’ 5’
T 5’ 3’ 3’ 5’
3’ 5’
T H C
3’ 5’

5’ 3’ 5’ 3’ 3’ 5’
T H C
A C G
3’ 5’ 3’ 5’ 5’ 3’

A.T G.C
 Guanine changes into Xanthin which pairs with Cytosine.
Xanthin can also pair with Thymine

5’ 3’
G
HNO2 5’ 3’
C 5’
3’ 5’ 3’ X
5’ 3’ X T
X 3’ 5’
C 5’ 3’ 3’ 5’
3’ 5’
C X T
3’ 5’

5’ 3’ 5’ 3’ 3’ 5’
G X T
C T A
3’ 5’ 3’ 5’ 5’ 3’

G.C A.T
 Nitrous acid produces exclusively
transition mutations
 Both C.G T.A & T.A C.G transitions
are produced
 Thus mutations can be reversed with the
nitrous acid
Hydroxl amine
 Specific base modifying mutagen which
adds a hydroxyl group to cytosine
producing hydroxlamine cytosine which
pairs with adenine instead of guanine
 This Leads to C.G T.A tranisitions
 Acts only on cytosine thus can not revert
the mutation produced
 Cytosine changes into hydroxlamine Cytosine which pairs
with Adenine instead of Guanine

5’ 3’
C
NH₂OH
G 5’hC 3’
3’ 5’ 5’ 3’
5’ 3’ hC A
hC 3’ 5’
G 5’hC 3’ 3’ 5’
3’ 5’
G A
3’ 5’

5’ 3’ 5’hC 3’ 3’ 5’
C A
G A T
3’ 5’ 3’ 5’ 5’ 3’

C.G T.A
Oxidative reactions:
 Reactive forms of oxygen like superoxide
radicals, hydrogen peroxide and hdroxyl
radicals produced in the course of normal
aerobic metabolism or by radiation, ozone,
peroxides, and certain drugs Cause
damage to DNA & induce mutations by
chemical changes
 Oxidation converts guanine into 8-oxy-7,8-
dihydrodeoxyguanine which mispairs with
adenine leading to G.C T.A
transversion
Intercalating agents
 Proflavin, acridine orange, ethidium bromide,
and dioxin
 They are about the same size as a nucleotide
 They produce mutations by sandwiching
themselves (intercalating) between adjacent
bases in DNA
 They distort the three-dimensional structure of
the helix and cause single-nucleotide insertions
and deletions in replication
 These insertions and deletions frequently
produce frameshift mutations
 Radiations:
Ionizing radiations:
In 1927, Herman Muller demonstrated that mutations
could be induced by X-rays.
X-rays, gamma rays, and cosmic rays are all capable of
penetrating tissues and damaging DNA.
 They remove electrons from the atoms that they
encounter, changing stable molecules into free radicals and
reactive ions which then alter the structures of bases and
break phosphodiester bonds in DNA.
 Ionizing radiation also frequently results in
double-strand breaks in DNA.
Mutation rates

 The frequency with which a gene changes

from the wild type to a mutant is reffered to
as the mutation rate.
 Expressed as the number of mutations per
biological unit i.e. mutations per cell division,
per gamete per round of replication

e.g. mutation rate for achondroplasia

(hereditary dwarfism) is about 4 mutations
per 100,000 gametes
 Mutation frequency:
Incidence of a specific type of mutation with
in a group of individual organism
e.g. for achondroplasia, the mutation
frequency in united states is about 2x10⁻⁴