5/17/2016

DNA MUTATION

BIO20 Introduction Prof. Ureah Thea A. Sevilla

to Biomimetics

MUTATIONS
Mutation is the alteration in the gene sequence.
 Hereditary (mutation on the germline)

 Change in structure of nucleotide sequence of the DNA

Mutation can be spontaneous or induced.

Mutation according to size:

 Gross Chromosomal mutation
 Point mutation

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Gross Chromosomal Mutation

 Down Syndrome

Gross Chromosomal Mutation

 Down Syndrome

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Gross Chromosomal Mutation

 Turner Syndrome

Gross Chromosomal Mutation

 Klinefelter Syndrome

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Point Mutation
A. Substitution mutation
single-base mutations, substitution of one or more
nucleotides by the same number of different
nucleotides.

Transition Transversion
purine to purine purine to pyrimidine (vice-versa)
A  G A  C G  C
pyrimidine to pyrimidine A  T G  T
C  T

Consequences of Substitution Point Mutation

SILENT MUTATION – no effect on the amino acid sequence of the protein.

Ex: CAG  CAA
Gln  Gln

MISSENSE MUTATION – results in the incorporation of a different amino acid

in the protein. The protein may be functional, partially functional or non-
functional.
CAG  CAC
Gln  His

NONSENSE MUTATION – results in the premature appearance of the stop

codon resulting to a shortened protein that is likely to be non-functional.
CAG  TAG
Gln  Stop

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Sickle Cell Anemia

Substitution Point Mutation

Given the following DNA:
Normal: 5’- ATG CCC AAA TGC GGG CGC – 3’
Mutant: 5’- ATG CCC AAA TAT GGG CGC – 3’

a. At what codon # did mutation occurred?

b. What type of base substitution?
c. What is the resulting AA of the mutated codon?
d. What is the consequence of the mutation?

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Substitution Point Mutation

Given the following DNA:
Normal: 5’- ATG CCC AAA TGC GGG CGC – 3’
Mutant: 5’- ATG CCC AAA TGA GGG CGC – 3’

a. At what codon # did mutation occurred?

b. What type of base substitution?
c. What is the resulting AA of the mutated codon?
d. What is the consequence of the mutation?

Point Mutation: Deletion and Insertion

Deletion Mutation – removal of one or more nucleotides in the
DNA.
Consequence of Deletion:
FRAMESHIFT MUTATION – a shift in the reading frame, which
produces a non-functional protein

ATT GGC AAA GCA TCA AAC TTA GGG CCC

deletion of G at
codon 4

ATT GGC AAA CAT CAA ACT TAG GGC CC…

--- reading frame was changed

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Point Mutation: Deletion and Insertion

Insertion – addition of one or more nucleotides in the

DNA

Consequence of Deletion:
FRAMESHIFT MUTATION – a shift in the reading
frame, which produces a non-functional protein

Deletion and Insertion Mutations are usually

caused by Replication Slippage

Frameshift Mutation
 Cystic Fibrosis is caused by a mutation in the gene
cystic fibrosis transmembrane conductance regulator
(CFTR).

F508, is a deletion at 508th position on the protein.

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Spontaneous Mutation
SPONTANEOUS MUTATION
likely mutations that occur because of the following:

 During DNA Replication

 Insertion or deletion during replication slippage

Induced Mutation
INDUCED MUTATION
are mutations that were acquired from the environment
(environmental factors).
 Physical Agents
- mutagens in the form of high energy radiation
 UV Radiation – leads to the formation of dimers which
can block DNA replication or interfere with base pairing.

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Induced Mutation: Physical Agents

UV Radiation

Induced Mutation
 Physical Agents
- mutagens in the form of high energy radiation
 X-rays
– leads to single and double-stranded DNA
breakage or may lead to formation of hydroxyl radicals
from water.

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Induced Mutation: Physical Agents

Ionizing Radiation: X-rays

Induced Mutation
 Chemical and Mutagens
 Base Analogues
-have similar structures to the DNA bases which
undergo tautomerism enabling to cause tautomer
mispairing
Example: 5-bromouracil

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Induced Mutation

 Base Analog: 5-
Bromouracil
5-BrU induces a point
mutation via base
substitution. This base
pair will change from
an A-T to a G-C or
from a G-C to an A-T
after a number of
replication cycles

Induced Mutation
 Alkylating agents
-adds a methyl or ethyl group to a base
- the largest class of potential mutagens present in man’s
environment

Example: N-nitrosamines (in cigarette smoke)

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Induced Mutation
 Alkylating agent: N-nitrosoamine
When N-nitrosamine is in the liver, it is metabolized by liver enzymes to
form alkylating agents which can attack guanine. If alkylation occurs, an
apurinic site is produced. During replication, an apurinic site may be
ignored resulting in a deletion in the daughter strand or a base selected at
random and place in the daughter strand.

Induced Mutation
 Deaminating agents
-removes amino groups from a base
Example: Sodium nitrite (used as preservative, color enhancer
and color fixative bacon, smoked fish, tocino, etc.)

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Induced Mutation
When ingested, NaNO2 is converted to nitrous acid in acidic
conditions. HNO2 removes functional groups from Adenine,
Guanine and Cytosine. Like deamination of adenine, will result in
the formation of hypoxanthine, a base analogue of guanine.

Induced Mutation
When ingested, NaNO2 is converted to nitrous acid in acidic
conditions. HNO2 removes functional groups from Adenine,
Guanine and Cytosine. Like deamination of adenine, will result in
the formation of hypoxanthine, a base analogue of guanine.

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Induced Mutation
 Intercalating agents
- contains a cyclic system that can interact with the bases of the
DNA
Examples:
Benzene (an organic solvent)
Benzo-a-pyrene (cigarette smoke, coal tar, automobile exhaust,
charbroiled food)
Aflatoxin (metabolic product of molds in peanuts, oils and grains)

Induced Mutations
Intercalating Agents
It physically binds to the bases by inserting itself between adjacent base pairs
because of its planar structure. This affects the opening of the DNA during
replication or transcription.

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Induced Mutations
Benzo(a)pyrene inserted
between base pairs

Induced Mutation
 Viral agents
some viruses contain oncogenes which can be
activiated once they have inserted their DNA in the
host’s genome.
When virus insert their DNA into the host’s genome,
the sequence of the bases of the host DNA may be
altered or certain destructive genes can be
activated.

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A Review of the Mutating Agents

Identify what type of mutagen are the following
whether physical, alkylating, deaminating, or
intercalating agents:
1. Sodium nitrite (NaNO2)
2. Benzene
3. UV-Radiation
4. N-Nitrosamine
5. Aflatoxin
6. X-Ray

References:
 Voet, Voet and Pratt, Principles of Biochemistry 3 rd
ed. Wiley Publication (2008).
 Lodish H, Berk A, Zipursky SL, et al. Molecular Cell
Biology, 5th edition. New York: W. H. Freeman;
(2004).
 Barnum, S., Biotechnology, Wadsworth – Thomson
Publishing Co. (1998).

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