GENETIC MUTATION

Asmarinah
Department of Medical Biology
Faculty of Medicine, University of Indonesia

Xeroderma pigmentosum patient

(extremely sensitive to sunlight)

DNA - Gene
The gene store genetic information encoded in the sequence
of nucleotide pairs in DNA

Inheritance is based on genes that are transmitted from parents

to offspring during reproduction with considerable accuracy

Mutation
Is the heritable changes in the genetic material.
The term mutation refers to
(1) change in the genetic material,
(2) the process by which the change occurs.
Mutation provides the raw material for evolution.
Without mutation, all of genes would exist in only one
form and alleles would not exist. Organism would not be
able to evolve and adapt to environment changes.

 Mutations range in size from a single DNA

building block (DNA base) to a large segment of a

chromosome.
An organism that exhibits a novel phenotype
resulting from a mutation is called a mutant.

* Mutation may occur in any cell and at any stage in the

development of a multicellular organism.
- germinal mutation occur in germ-line cell and will
be transmitted to progeny
This type of mutation is present throughout a persons life in
virtually every cell in the body.

- somatic mutation occur in somatic cell, occur in the

DNA of individual cells at some
time during a persons life, and will
not be transmitted to the progeny.

Single-site or point mutation within individual gene

be classified:
- Substitution
In which one or more base-pairs are substituted with
other base pairs
- Deletion
In which one or more base-pairs are lost
- Insertion
In which one or more base-pairs are inserted into
sequence

Types of point mutation:

The mutation of a wild-type gene to a form that result in

a mutant phenotype forward mutation.
When a second mutation restores
phenotype,
the process is called reverse mutation.

the

original

Reverse mutation may occur in two different ways,

1) by back mutation, a second mutation at the same site
in the gene as the original mutation, restoring the
wild type nucleotide sequence,
2) by suppressor mutation, a second mutation at a
different location in the genome, which compensates
for the effects of the first mutation.

The deletion and insertion of one or two base-pairs

within the coding sequence of a gene will alter the
codon reading frame (i.e the series of base triplets that
specify amino acid during translation) in mRNA.
frameshift mutations.

Mutation can alter the sequences of nucleotide pairs in

genes, that cause changes in the amino acid sequences of
the polypeptides missense mutation.
Mutation that alter the sequences of nucleotide pairs in
genes, but cant cause changes in the amino acid
sequences of the polypeptides silent mutation
If the point mutation of base-pairs produce a stop codon
(UAA, UAG and UGA) nonsense mutation

Many mutation have no effect on the phenotype of

organism neutral mutation.
Most mutation with phenotypically recognizable
effects result in decreased gene-product activity or no
gene-product activity.

Mutation process and the expression of wildtype and mutant alles

Recessive mutant alleles often result in blocks in

metabolic pathway
For example: 5 disorders are caused by autosomal
recessive mutation with defect in phenylalaninetyrosine metabolism:
-Phenylketonuria
-Tyrosinosis
-Tyrosinemia
-Alkaptonuria
-Albinism

Based on the cause of the mutation, there are:

* Spontaneous mutation
Occur infrequently, without a known cause.
For eukaryotes, mutation rates range from about
10-7 to 10-9 per nucleotide pair per generation
Mutation rate per gene varies from about 10-4 to
10-7 per generation
* Induced mutation
Resulting from exposure of organism to physical
and chemical agents, called mutagens

Mutagenic agent (Mutagen)

result in induced mutations. It is classified into:
1. Chemical
- alkylating agent, that transfer alkyl group (CH3-,
CH3CH2, and so forth) to the bases in DNA
Example: Mustard gas [Di-(2-chloroethyl) sulfide]
EMS (Ethyl methane sulfonate)
- Basa analogs, that have similar structure to the normal
base and are incorporated into DNA during
replication
Example: 5-Bromouracil (BU)
2-Aminopurin (2-AP)

- Deaminating agent that deaminate the

amino groups in bases
Example: Nitrous acid (HNO2)
- Hydroxylating agent, that hydroxylase amino
groups in the bases caused transition mutation.
Example: Hydroxylamine
- Acridin dyes, that intercalate DNA molecule, so
doing, they increase the rigidity and alter the
conformation of the double helix
Example: Proflavin
Acridine orange

Some potent chemical mutagens

2. Physical agents
- Ionizing radiation
High-energy rays collide with atom and cause the
release of electron, creating free radical or ion. It
also induces gross changes in chromosome
structure
Examples: X-rays, gamma rays and cosmic rays
- Nonionizing radiation
Lower-energy that penetrate only the surface layer
of cells and dont cause ionizations.
Example: UV light cause pyrimidine hydrate
pyrimidine dimer

3. Transposable Genetic
Elements (transposons)

DNA element that can

move from site in the
genome to another site.
The insertion of a
transposons will often
render the gene
nonfunctional.

Three major factors for the occurring of mutation :

1. The accuracy of the DNA replication machinery
2. The efficiency of the mechanisms that have evolved
for the repair of damaged DNA.
3. The degree of the exposure to mutagenic agents
present in the environment.

The Mechanism of Mutation

1. Tautomerisation in DNA replication process
Tautomerisation is a process in which hydrogen
atoms moves from one position to another position
in a purine or pyramidine .
Mutation resulting from tautomeric shifts cause:
- Transition mutation: replacement of purine with
other purine or of pyrimidine to other pyrimidine
- Transversion mutation: replacement of purine with
pyrimidine and vice versa

Tautomeric forms of the four

common bases in DNA

The shift of hydrogen atoms

between the number 3 and 4
position of pyrimidine and
between the number 1 and 6
position of the purine change
their base-pairing potential

Mechanism by which tautomeric shift in the bases in DNA

cause mutations

2. Failure in DNA repair process

DNA repair mechanisms are classified into:
1. Excision repair
Consist of 3 steps:
a. DNA glycosylase enzyme
recognized and then excises the
damage bases in DNA
b. DNA polymerase fills nucleotide
in the gap
c. DNA ligase seals the break to
complete the repair process

- Light-dependent repair
(photoreactivation)
DNA photolyase
recognizes and binds to
thymine dimmers, and then
uses light energy to cleave
cross-link

Inherited human disease with DNA mutation

(examples)
-Sickle cell anemia :
Pauling et al., (1949) discovered: substitution of
nucleotide A to T that substitute amino acid glutamic
acid in the normally hemoglobin (Hb A) to valine
(defective hemoglobin (Hb S).
introduced molecular disease
term
set the foundation of molecular
diagnosis

Inherited human disease with defects in DNA repair

Xeroderma Pigmentosum
It caused by defect in the repair of UV-induced damaged
to DNA

High frequency
to skin cancer

Mutation is one of sources for polymorphism

Polymorphism
many form : a set of two or more alternative of
normal phenotypes
Genetic polymorphism:
The occurrence of two or more allele in a locus in
which its frequency is more than 1% in
population. Example: ABO and MN blood group
If the alleles with frequencies of less than 1% are
called rare variants . Example: O Bombay blood
group

 Polymorphisms are responsible for many of the

normal differences between people such as eye color,
hair color, and blood type.
Although many polymorphisms have no negative
effects on a persons health, some of these variations
may influence the risk of developing certain disorders.

Forms of genetic polymophism

1. Single Nucleotide Polymorphism (SNP), a
polymorphism in DNA sequence consisting of
variation in a single base.
Example: Missence mutation that caused by a
nucleotide substitution

2. Variable number of tandem repeats (VNTR), a type

of DNA polymorphism created by a tandem

arrangement of multiple copies of short DNA
sequences.

3. Micro satellites (Short Tandem Repeat Polymorphism =

STRP)
A polymorphic locus consisting of a variable number of
tandem repeated bi-, tri-, or tetra nucleotide units such as
(TG)n, (CAA)n, or (GATA)n.
Different numbers of units constitute the different alleles

Gene polymorphism
Occur in 1 in 1000 DNA bp in human genome
over 3 million SNPs in the entire human genome
Is reflected in the diversity of the gene products, such
as structural protein, enzymes, channel proteins,
receptors, etc.

Use of polymorphism
Medical forensic individual characteristic
Disease susceptibility, influence drug response
Genetic relationship (within & between population)
molecular evolution

Genetic Mutation/Polymorphism
can be detected by following techniques:
* RFLP (Restriction Fragment Length Polymorphism)
* PCR (Polymerase Chains Reaction) RFLP
* PCR DNA Sequencing
* Southern Blot

PCR-RFLP method
(for diagnosis of sickle cell anemia)

1300
1100

PCR-DNA sequencing method:

Mutation in exon 6 of VDAC3 gene in sperm with low motility
174
Sequence of PCR
product from sperm
with normal motility
Posisition 174 : AAG
(Lysine)

Sequence of PCR
product from sperm
with low motility
Posisition 174 : GAG
(glutamic acid)

References:
Alberts et al., 2008. Molecular Biology of the Cell. 5th ed.
Snustad & Simmons. 2010. Principles of Genetics . 5td ed.
Karp. 2005. Cell and Molecular Biology. 4th ed.
Nussbaum et al. 2001. Thompson & Thompson Genetic
Medicine. 6th ed.