Genetics & Molecular Biology

Mutation
Definition: Mutation may be defined as a sudden heritable change in the characteristic of an
usually
organism that involves an alteration in the DNA base pair or chromosome. Mutations are
random, recessive, deleterious in effect and recurrent in occurrence.
" The tem "mutation" was coined by Hugo de Vries (in 1880's) who observed phenotypic
changes in Oenothera lamarckiana that resulted due lo mulation, However, research on mutation
started in 1910, when Morgan worked on Drosophila and reported mutated eye colour.
Types of Mutation:
According to their site of occurrence, imulations may be of the following types:
" Chromosomal mutation: If mutation involves a gross change hat affects structure or number of
chromosomes, then it is known as chromosomal mulation.
" Point mutation: Mutation that involves a change in a single base pair in the DNA molecule is
known as point mutation. Point mutation may occur in the coding (gene mutation) or noncoding
region of the DNA.
Gene mutation: It is a type of point mutation that alters the function of a single gene.
On the basis of their consequence, mutations may be of the following types:
" Silent mutation: A silenl mutation is a lype of point mutation that changes one codon to another
codon in the mRNA, resulting in no change in the amino acid sequence or function of the
protein.
Ncutrl mutation: Ancutral mutalion is an alteration in base-pair of a gene that changes the
codon in the mRNA such that the resulting amino acid is ch:amicaly equivalent and produces no
detectable change in he protein function.
" Missense mutation: Apoint mutation that changes one codon to another in the mRNA so that it
specifies a different amino acid than the one specified by the original codon is known as
missense mutation. This results in a change in the protein function.
Nonsense mutation: Apoint mutation in a gene that changes an amino-acid-coding codon in the
mRNA toa stop codon is known as nonsense mutation.
Frameshift mutation: A mutation that leads to the insertion or deletion of one or nore base pairs
in a gene that shifts the mRNA reading frame in all codons after the mutational site.
Mutation Mutatlon Mutation Mulatlon
Normal gene Addiion

DNA

GUU UAU GUA UGG yAG

Pro Glu Glu Cys Pro Arg Gly Val
NA UUU UUC CUU

retela Phe Phe Leu Val Tyr Val Tip Stop

Silent Neutral Missense Nonsense Frameshift

Based on the tissue of origin, mutations are of the following types:

Somatic mutations: When nutation occurs in a somatic cell (in multicellular organisms), that do
not produce gamete, is known as somatic mutation. In somatic mutation, the mutant
characteristic affects only Lhe individual in which (he mutation occurs and is not passed on to the
succeeding generation.
Germ-line mutation: í mutatiuns arise in cells that prod::ce gamete, then it is known as germ
line mutation. A germ-line mulation may be transmitted to the next generation, producing an
individual with the mutation in both its somalic and germ-lir: cells.

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(Genetics & Molecular Biology
Based on the causes of mutations, it may be of the following lypes:
" Spontancous mutation: Mutation hat is nol induced by a chemlenl or physical agent bul arien
Irom natural changes in DNA structure or eplication errors is known os spontaneoun mulation.
" Induced mutation: Any mutation thal resulls (rom the influence of a chemlcal or pl1ysical agent
(mutagen) is known as induced mulation.
Based on the direction of mutatlons, il may be of following types:
" Forward mutation: Apoint mutatlon in awild-type allele that changes It to a mutant allele is
known as forward nutation.
" Reverse mutation: A point mutation in a mutant allele that changes it back to the wild-lype
allele is known as reverse mutation.
This reversion can result in wild-type amino acid (lrue reversion) or to some other amino acid
(partial reversion) thus a complete or partial function may be restored.
" Suppressor mutation: Effects of a mutalion may be diminished or abolished by a second
mutation at a different site from the original mutation known as the suppressor mutation.
Suppressor mulations that occur within the same gene where the original mulation ocurred,
but at a different site and are known as intragenic suppresors, or they may occur in a different
gene and are called intergenic suppressors.
Based on ils effect on gene function, mutation may be of the following types:
Loss-of-function mutation: A mutation that result that inactivates normal gene function is
known as loss of function mutation. They qr gengrally reccssive.
" Gain-of-function mutation: Mutafog uhaf resul it gchyation of apreviously inactive gene is
known as gain of function mutamag. They aregenerallpdòminant.
Additionally point mutation can be syoupedint vogeneralcptegories: base-pair substitution and
base-pair insertion or deletion. Transitions Possible
base chauges
Base-pair substitution mutationA typal of mutatio that
involves a change of one base patoanother. They. areo2 types: Purine Purine

A. Transition mutation: It is atype de fmrgtignyaWhich apurine

is substituted for a purine, or a pyrimidine substituted for a
Pyrimidine. Pyrimidine Pyrimldine
Transversions
B. Transversion mutation: In this type of mutation a purine is
substituted for a pyrimidine or vice versa.
Purlne Pyrimidine
" Base-pair insertion or deletion: It is a type of mutation that
involves an insertion or deletion of a base pair of DNA and leads
A
to frameshift mutation. Pyrimidine Purine

Mutagens: Any physical or chemical agents that induces mutation are known as mutagens.
Mutations are can be artificialy induced at relatively higher frequencies by treatment with radiations
(physical mutagens) or chemicals (chemical mutagens). Mutagens may be grouped as follows:
Carcinogens: Any chemical that causes neoplastic transformation of cells (cancer).
Clastogens: Any chemical that causes fragmentàtion of chromosomes.
Teratogens: Any chemical that causes developmental abnormalities.
Types of Mutagens:
A. Physical Mutagen:
Non-ionizing radiations: UV-radiation (most effective at 260 nm)
Ionizing radiations: Particulate: a-rays, B-rays and thermal neutrons.
Non particulate: X-rays, y-rays.
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Genetics & Molecu lar Biology
B. Chemical Mutagen:
Alkylating agents: Ethyl methane sulphonate (EMS), Methyl methane sulphonate (MMS).
Base analogues : 5-bromo uracil, 5-chloro uracil.
Intercalating agents: Proflavin, Acridine orange
" Base-modifying
agents: Nitrous acid, Hydroxyl amine, Sodium azide.
Molecular Basis of Mutation:
A. Toutomeric Shifts (Tautomerisation):
Cownon fonns Rare foms ANdard basesalring arraemeats
" After DNA was identified as Proton shin
the genetic material, it was
pointed out that structures of
Thymine Adenine (comnon formi
Tymine (common lorm
bases in DNA are not static
and recognized that purines
and pyrimidines found in Guanine
DNA exist in alternate Cyyosiae (common form) Cuanioe (common lormb
Anomalous basepaleing arrangnents
chemical forms called
structural isomers Cnosine

tautomers, each differing by

Adenine (commam lorm)
only a single proton shift in w Cyoane (rare forn)

the molecule. Adentne

" Such a shift can change the

bonding structure of the base Tautomeric Shifts Thymine tcommon form Cuae rare forrnl

and may result in base pair Tautomnerization to

change or mutations (spontaneous mutations). rare imíno form
" In DNA, the keto form is responsible for the normal Watson-Crick
base pairing of T=A and C=G while, non-Watson-Crick base
G

pairing can result if a base is in a rare tautomeric state, the enol T C

Transitlon
form. When the bases are present in their rare enol forms, A mutatlon

anomalous A = C and G=Tbase pairs can result but the pairing is No

mutation T
always between a purine and pyrimidine.
The effect leading to mutation occurs during DNA replication when a rare tautomer in tenplate
strand matches with a non-complementary base. In next round of replication, the 'mismatched'
base pair is separated and each specifies its normal complementary base resulting in mutation.
B. Base Analogs:
OH
" Base analogues are mutagenic chemicals
that have molecular structures similar to
H
H
the nitrogenous bases present in DNA.
" So they are incorporated into DNA in Thymine S-Bromouracil (kelo form) 5-Bromouracil (enol form)
place of the normal bases during
replication. They increase the frequency
of mis-pairing and therefore mutation.
" One base analog is 5-bromouracil (5BU),
which has a bromine residue instead of H H
S-BU (keto form) Adenlie S-BU (enol form) Guanine

the methyl group of thymine. In its

it pairs
normal state, 5BUresembles thymine and pairs with adenine in DNA but in its rare state,
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Genetics & Molecular Biology
with guanine. If 5-BU is incorporated into DNA in place of thymine, and a tautomeric shift to its
rare enol form results in A =Tto G = C
transition after one round of replication. 3
DNA
" 2-amino purine (2-AP) is another base analog replicallon
A:5B0
of adenine that is mutagenic. In addition to its DNA
base-pairing afinity with thymine, 2-AP can replication
G:SBU
also base-pair with cytosine, leading to
possible transitions from A = T to G C G'orA:58U

following replication.
C. Alkylating Agents:
Alkylating agents are chemicals that donate an alkyl group, such as CHs (methyl) or CHs-CH2 (ethyl),
Type of
to amino or keto group of Original base Mutagen Modifled base Pairing partner mutation
nucleotide bases.
" For example, ethyl H. CHy

methane sulfonate (EMS) G -H

EMS
CGTA

adds an ethyl grOup to

Alkylation
guanine, producing 06
ethylguanine, which pairs Guanine 06-Ethylguanine Thymine
with thymine. Thus, EMS (ethyiguanine)
produces CGtoT=A transitions affer DINAHeplfcatio, EMS is Alkylation A
also capable of adding an ethyl goup tothymine, prodycing 4 (EMS)
ethylthymine, which then pairs Witt guanie edding toa ]= A T
Anomalous
T
to C =G transition. Because EM$ roducesboth GtßT A base pair
Transitlon
and T- Ato C=G transitions, mtations proguced by EM can c mutation

be reversed by additional treatmet With EMS.

Methyl methane sulfonate (MMS)ustard
Dan gs boActs as
alkylating agents.
D. Deaminating Agents: Type of
Nitrous acid (HNO:) is a Original base Mutagen Modified base Pairing partner mutation
very potent deaminating NH, O****H-N
agent that acts directly
Nitrous acid
on DNA by oxidative CGTA
(HNO,) H
deamination of the bases Deamination H
that contain amino Cytosine Uracil Adenine

groups- adenine, cytosine, and guanine. In the process of

deamination, an amino group is converted to a keto grOup.
" Treatment of cytosine with nitrous acid produces uracil, which G
pairs with adenine to produce, a CG to T=A transition A
C
mutation during replication. A
Treatment of guanine with nitrous acid produces xanthine, but Deaminatlon T
(HNO2)
because this purine base has the same pairing properties as U* Transition
mutation
guanine, no mutation results.
Nitrous acid modifies adenine to produce hypoxanthine, a base that pairs with cytosine rather
than thymine, which results in an A =T to G=C transition mutation.

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Genetics & Molecular Biology
Mutatlon by addillon
E. Intercalating agents: Molecde of
ntercalatng agent
Chemical agents such as proflavin, acridine, and fomplale DNA sNrand 6 ATOAGTIACT 3
5'
Now DNA strnnd STAOTGGAATOA
ethidium bronnide (used to stain DNA in gel 0G8 nm-
-A randomly choen
baso in lnserled opposte
electrophorsis)-insert (intercalate) themselves or
Inlercalaling agent, hore,
tho base is G
"sandwich themselves between the stacked base pairs in Subsequent replication
DNA" in one or both strands of the DNAdouble helix,
of now siand

causing the helix to relax. 5 ATCAGCTTACT 3

TAGTGGAA TGA
If the intercalating agent inserts itself between adjacent Rosul: Iromeshin mutation
due lo insortlon ol
base pairs of the DNA strand that is the template in one bas pair (CG)

DNA replication, an extra base (chosen at random, G in

the figure) is inserted into the new DNA strand opposite Mutation by deletl on
to the intercalaing agernt. After one more round of Template DNA Strand
New DNA strand
5'
3'
ATCAGTTACT
TAGTC ATGA 5
replication, during which the intercalating agent is lost, Intercalaling
the overall result is a base-pair addition mutation (C-G agenl
Aeplication ol new slrand after
is added). Intercalattng agent lost
" If the intercalating agent inserts itself into the new DNA
strand in place of a base, then when that DNA replicates
3
ATCAQTACT
TAGTCATGA
after the intercalating agent is lost, the result is a base
pair deletion mutation (T = A is lost).
F. Dimerization: Ultraviolet Radiation
UV light
" UV ray do not possess sufficient energy to
induce ionization, but they are readily absorbed TCCAACGTAG
GOGCAATC
by purine and pyrimidine bases. Thymine Covalent4
bases bonds
Because of their lower energy, they penetrate 5
tissues only slightly, usually the surface layers of Sugar-phosphate
cells in multicelluler organisms. The maximum backbone

absorbtion of UV by DNA is at a wave length of

254 nm. 18.22 Pyrimldine dimers result from ultraviolet light.
(a) Formation of thymine dimer. (b) Distorted DNA
One effect of UV radiation on DNA is the
formation of abnormal chemical bonds between adjacent pyrimidine molecules in the same strand
of the double helix.
This bonding is induced mostly between adjacent thymines, forming peculiar structures called
thymine dimers, usually designated as T^T.
" This unusual pairing produces a bulge in the DNA strand and disrupts the normal pairing of T
with corresponding adenine bases on the opposite strand.
Replication cannot proceed past the lesion, so the cell will die if enough pyrimidine dimers remain
unrepaired.

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