MUTAGENT &

DNA MUTATION

• by: Puspa Julistia Puspita, M.Sc. @Genetika dan Biologi Molekular

 MUTAGEN
Is an agent that can changes the genetic material,
usually DNA, of an organism and thus increases the
frequency of mutations above the natural background level.

Mutagen does not create cancerous

cells, while Carcinogen does and can
often lead to cancer. Mutagen only
causes mutation, but is usually mixed
up with Carcinogen due to its
characteristics that ultimately lead to
cancerous cells.
 TYPE OF MUTAGENT
CHEMICAL- • Base analogue (which can substitute for
MUTAGENT DNA bases during replication and cause
transition mutations)
• Intercalating agent (molecules that may
insert between bases in DNA, causing
frameshift mutation during replication)
• Alkylating agent
• Hydoxylating agent
• Deaminating agent
• Transposable element?
PHOTO- Ionizing radiation (X-, -rays)
MUTAGENT UV radiation

BIOMUTAGENT Bacteria
Virus
Human?
 BASE ANALOGUE
● 2AP

● 5BU
 5-BROMOURACIL INTERACTIONS

Guanine
Adenine Thymine 5-Bromouracil (keto form) 5-Bromouracil (enol form)

Deoxyribose Deoxyribose
Adenine 5BU (normal keto state)

Deoxyribose
Deoxyribose
Guanine 5BU (rare enol state)
 EFFECT OF BASE ANALOGUE (5 BU)
● 5BU ← T ● 5BU ← C

A. G.
T C

A. A.
BU BU

A. G. G. A.
+ +
T BU C BU

A. A. G. A. G. G. A. A.
+ + + +
T T C BU C C T BU

Error in replication A.T  G.C Error in replication G.C  A.T

INTERCALATING AGENT (INTERCALATOR)
 EFFECT OF INTERCALATOR
Intercalator: acridine orange

A.T A.T
A.T
T.A A.T
C.G T.A
C.G

TYR Glu Thr Gly Ile

5’ T A C G A A T C G G G T A T T 3’
3’ A T G C T T A G C C C A T A A 5’
Replication in the presence of intercalator
5’ T A C G A G A T C G G G T A T T 3’
3’ A T G C T C T A G C C C A T A A 5’
TYR Glu Ile Gly Tyr
ALKYLATING AGENT
 ALKYLATING AGENT
● Nitrosoguanidine CH3 CH2-CH3
O N N O N N
CH3 CH2-CH3
Dimethyl nitrosamine Diethyl nitrosamine
CH3
● N-methyl-N’-nitro-N-nitrosoguanidine O N N
H
C N
NH NO2
Alkylating agent
O
N Pairs normally
NH with cytosine
CH3
N N NH2
O
Guanine
N NH
Sometimes pairs with thymine
N N NH2
O6-methylguanine
DEPURINATION (Hydroxylating agent)

Terlepasnya ikatan
glikosidik antara basa
purin dan gula
deoksiribosa
 EFFECT OF DEPURINATION

G.C

G.C

G.C
Depurination
G.C .C Replication
Pu-Py pair
.A
Efeknya : mutasi (pergantian
basa) acak basa baru .C
diadakan secara acak jadi
tidak sama dengan basa T.A
mula-mula Py-Pu pair
 DEAMINATION OF VARIOUS BASES
Gugus asam amino tersingkir dari basa nitrogen
DEAMINATION BY HNO2

Nitrous acid

HNO2

O
N
OH
 EFFECT OF DEAMINATION

HNO2 Deoxyribose
Deoxyribose
Adenine Hypoxanthine Cytosine

HNO2
Deoxyribose Deoxyribose
Uracyl Adenine
Cytosine

HNO2
Deoxyribose Deoxyribose
Guanine Cytosine
Xanthine
TRANSPOSABLE ELEMENT

Transposom adalah urutan

DNA yang dapat menghapus
atau memasukkan dirinya ke
dalam genom.

Transposom berpindah dari satu

bagian ke bagian lain dari
DNA, cenderung membuat
beberapa perubahan dalam
genom sehingga menimbulkan
mutasi dan perubahan genom
 BIOMUTAGENT

 Bacteria (occur in bacteria, plant)

 Virus (occur in bacteria, plant, and mamal)
 Human ?
GENE TRANSFER
 HORIZONTAL GENE TRANSFER

• Two types of cells:

1.Donor: transfers DNA to recipient
2.Recipient: receives the DNA

• Three methods of horizontal gene transfer:

1. Transformation
2. Transduction
3. Conjugation
 TRANSFORMATION

• Donor cell wall

can rupture, and
DNA that is
normally tightly
packed can break
up into pieces and
explode through
cell wall

• These pieces can

then pass
through cell walls
and cytoplasmic
membranes of
recipient cells and
get integrated
into their
chromosomes
 TRANSDUCTION
• DNA is transferred from one bacteria to another by virus that infects bacteria
(bacteriophage)
 CONJUGATION (1)
• Transfer of DNA by contact of two
bacterial cells
• Can transfer plasmid or chromosome
• Two types of cells:
1.Donor cell- contains
fertility plasmid and is
designated F+
2.Recipient cell- does not
contain fertility plasmid and
is designated F-
• DNA transferred from F+ to F-
 CONJUGATION (2)
• In some cells carrying F factors, the F factor integrates into the host
chromosome
• Now called Hfr cell

• Conjugation between Hfr

and F-
– Chromosome
replicates
– Transferred to F- cell
– Usually chromosome
breaks off before
completely
transferred
– Generally remains F-
because does not
receive F factor
 R PLASMID

• Resistance
plasmid-
confer
antibiotic
resistance

• Two parts:
1. Resistance
genes (R
genes)
2. Resistance
transfer factor
(RTF)
MUTATION
 MUTATION

 Definition of Mutation: change/defect on DNA (nucleic acid)

 The Cause of Mutation:
 Random thermal collison
 Error in replication
 Mutagen
 Effect of Mutation
 Silent  no effect on the phenotype
 Repression/activation of genetic expression
 amount of protein, activation of active protein
 Change in base sequence  change in amino acid sequence of protein
 Rate of Mutation
 Rate of change/defect Vs rate of repair
 1 mutation/107 base pairs pergeneration
 mutational hot/rare spot  ± 25x higher/lesser probability
 Mutagen
 Mutational-inducing agent
 Various types and mechanisms
 Type of Mutation
 Based on the location
 Regulatory mutation
 Structural mutation
 Mixed (regulatory and structural) mutation
 Based on the change/defect
 Base-structural mutation
 depurination
 deamination
 dimerisation
 etc.
 Base-sequence mutation
 point mutation
 substitution
 insertion/addition
 deletion
 oligonucleotide mutation
(chromosomal mutation)
 translocation
 invertion substitution
 duplication
 insertion
 deletion
Example effect of mutation
A nonsense mutation and
its effect on translation
 EXAMPLE OF BASE-SEQUENCE MUTATION

Wild type (Original sequence)

A T G A C C A G G T C

Base substitution Base rearrangement

From C From CCAG
A T G A C T A G G T C A T G A G A C C G T C

Base addition Base deletion

Addition of A Missing C
A T G A C A C A G G T C A T G A CA G G T C
 EXAMPLE OF BASE-STRUCTURAL MUTATION
Oxydation, hydrolysis or methylation

 = uncontrolled methylation;  = oxidative damaged;  = hydrolytic reaction

(deamination/depurination)
Dimer formation O O
H H
P C N P C N
N C O N C O
C C C C
H CH3 UV radiation H CH3
O H O H
P C N P C N
N C O N C O
C C C C
H CH3 H CH3
adjacent thymine bases thymine dimer
EFFECT OF BASE SEQUENCE MUTATION (ADDITION) ON READING FRAME

Original sequence

One nucleotide
added

2nd nucleotide First frame shift

added

3rd nucleotide
added Second frame shift

New amino acid inserted Restored to original sequence

 BASE SEQUENCE MUTATION: VARIOUS POINT MUTATIONS
(-) strand T substituted for C
mRNA Transcription
Translation
Base substitution
Missense mutation results in
an altered amino acid residue
Original sequence C
(-) strand Add a base
mRNA Transcription Insertion Transcription
Translation Translation

Frameshift results in scrambling

of downstream mRNA sequence
Deletion and a new sequence of amino acid
C Delete a base

Transcription
Translation

Deletion results in scrambling of downstream

mRNA sequence and a new sequence of amino acid
 EXAMPLE OF BASE SEQUENCE (POINT) MUTATION

 Point mutation on E. coli tryptophan synthetase structural gene

DNA for E. coli Positions of mutations

tryptophan 5’ 3’
synthetase gene

Positions of
mutations

Normal amino
acidsynthesized

Altered amino
acid resulting
from mutation
CHROMOSOMAL MUTATION
 CHROMOSOMAL NUMBER MUTATION

Euploid Variation in the the fundamental

number of chromosome (genom).

Autopoliploid Amplifikasi number of chromosome

of the same species

Alopoliploid Amplifikasi number of chromosome

of different species

Aneuploid Variation of chromosomal number

due to substraction or addition of one
or small amount chromosome as
results of failure during separation
 DOWN SYNDROME

Threesomy chromosome 21
→ (total) 47 chromosome
• Ditemukan oleh Longdon Down tahun 1866
• Ciri-ciri
- Kariotipe 47, XX atau 47 XY
- Telapak tebal spt telapak kera
- Mta sipit miring ke samping
- Bibir tebal, lidah menjulur, liur selalu
menetes
- Gigi kecil dan jarang
- Iq rendah
 KLINEFELTER SYNDROME

Extrachromosome X in
boy → XXY → (total)
47 chromosomes
• Ditemukan pada tahun 1942
• Ciri-ciri
- Kariotite 47, XXY diderita oleh pria
- Bulu badan tidak tumbuh
- Testis mengecil, mandul
- Buah dada membesar
- Tinggi badan berlebih
- Jika kromosom X lebih dari dua mengalami
TURNER SYNDROME

X monosomy in girl → XO
→ (total) 45 chromosomes
• Ditemukan oleh H.H Turner tahun 1938
• Ciri-ciri
• Kariotipe 45 XO (44 autosom + satu
kromosom X)
• Tinggi badan cenderung pendek
• Alat kelamin terlambat perkembangannya
• Sisi leher tumbuh tambahan daging
• Bentuk kaki X
• Keterbelakangan mental
 CRI DU CHAT SYNDROME
Deletion of chromosome no. 5 → retarded mental, small head, crying
like a cat, often die during infant

Baby with
Cri du chat
syndrome

Same boy
after 4
years old