GENETIC MUTATION

Asmarinah

Department of Medical Biology

Faculty of Medicine, University of Indonesia
 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 exis. Organism would not be
able to evolve and adapt to environment changes.
* 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
- somatic mutation  occur in somatic cell, occur only in
descendant of that cell and will not
be transmitted to the progeny.

An organism that exhibits a novel phenotype resulting

from a mutation is called a mutant.
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 the original phenotype,

the process is called reverse mutation.

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 can’t 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 wild-
type 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 phenylalanine-
tyrosin metabolisme:
-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 electrone, 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 don’t 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 insersion 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 pyrimidine .
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
 The effects of tautomeric shift in nucleotide

b. 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 glycosilase 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 dimers, and then
uses light energy to cleave
cross-link
Inherited human disease with DNA mutation

- “Sickle cell anemia”: substitution of nucleotide A

to T that substitute amino acid glutamic acid in the
normaly hemoglobin (Hb A) to valine (defective
hemoglobin (Hb S).

- “Tay-Sachs disease”: mutation in the HEXA gen

which encodes the enzyme hexosamidase A to
convert ganglioside GM2 into GM3 in nerve cells.
In the absence of the enzyme, ganglioside GM2
accumulates and blocks neuron activity.
Inherited human disease with defects in DNA repair
•Xeroderma Pigmentosum
extremely sensitive to sunlight. It caused by defect in
the repair of UV-induced damaged to DNA
•Cockayne syndrome
Defect in nucleotide excision repair
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
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 polymophism created by a tandem
arrangment of multiple copies of short DNA
sequences.
3. Microsatellites (Short Tandem Repeat Polymorphism =
STRP)
A polymorphic locus consisting of a variable number of
tandemly repeated bi-, tri-, or tetranucleotide 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: - predisposition
- tendency vs. resistence
- advantage vs. disadvantage
•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
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., 2002. Molecular Biology of the Cell. 4th ed.
•Karp. 2005. Cell and Molecular Biology. 4th ed.
•Nussbaum et al. 2001. Thompson & Thompson Genetic
Medicine. 6th ed.
•Snustad & Simmons. 2003. Principles of Genetics . 3rd ed.