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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
 

Gross Chromosomal Mutation

   
 
  • Down Syndrome

 Down Syndrome
Gross Chromosomal Mutation  Down Syndrome

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Gross Chromosomal Mutation

   
 
  • Turner Syndrome

 Turner Syndrome
Gross Chromosomal Mutation  Turner Syndrome
Gross Chromosomal Mutation  Turner Syndrome
Gross Chromosomal Mutation Klinefelter 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
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).

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

Induced Mutation INDUCED MUTATION are mutations that were acquired from the environment (environmental factors).  Physical

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Induced Mutation: Physical Agents

   
 

UV Radiation

Induced Mutation: Physical Agents UV Radiation
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.

 
Induced Mutation  Physical Agents - mutagens in the form of high energy radiation  X-rays
Induced Mutation  Physical Agents - mutagens in the form of high energy radiation  X-rays

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Induced Mutation: Physical Agents Ionizing Radiation: X-rays
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

 
Induced Mutation  Chemical and Mutagens  Base Analogues -have similar structures to the DNA bases

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Induced Mutation  Base Analog: 5- Bromouracil 5-BrU induces a point mutation via base substitution. This
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)

 
Induced Mutation  Alkylating agents -adds a methyl or ethyl group to a base - the

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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  Alkylating agent: N-nitrosoamine When N-nitrosamine is in the liver, it is metabolized by
 

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.)

 
Induced Mutation  Deaminating agents -removes amino groups from a base Example: Sodium nitrite (used as
Induced Mutation  Deaminating agents -removes amino groups from a base Example: Sodium nitrite (used as

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Induced Mutation

   
 

When ingested, NaNO 2 is converted to nitrous acid in acidic conditions. HNO 2 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, NaNO is converted to nitrous acid in acidic conditions. HNO removes functional
Induced Mutation When ingested, NaNO 2 is converted to nitrous acid in acidic conditions. HNO 2
Induced Mutation
When ingested, NaNO 2 is converted to nitrous acid in acidic
conditions. HNO 2 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

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 Mutation  Intercalating agents - contains a cyclic system that can interact with the bases
Induced Mutation  Intercalating agents - contains a cyclic system that can interact with the bases
Induced Mutation  Intercalating agents - contains a cyclic system that can interact with the bases
Induced Mutations Intercalating Agents It physically binds to the bases by inserting itself between adjacent base
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 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 (NaNO 2 )

  • 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, 5 th edition. New York: W. H. Freeman;

(2004).

  • Barnum, S., Biotechnology, Wadsworth Thomson Publishing Co. (1998).