Mutation & Mutagenesis

Muhammad Dawood
 The Problem
• DNA extremely long, fragile
• Subject to both physical and chemical damage
• Consequences could be lethal for organism or
offspring
 Mutation
• A heritable change in the base sequence of
DNA
• Point mutation- change in a single base
position
– Additions
– Deletions } Frameshift mutations
– Substitutions
• Transitions
• Transversions
• Multiple mutations
 Consequences of Mutation
• Silent Mutation---base change, no amino acid change
• Neutral Mutation--- Base change resulting in aa
change that does not affect protein function
– EX. Apartic acid (D) Glutamic acid (E)
• Missense mutation---altered codon, new aa with
different chemical properties. Function affected.
• Nonsense mutation---base pair substitution results in
a stop codon (and shorter polypeptide)
• Frameshift mutations—additions or deletions. Peptide
may be longer or shorter.
 Other Terms
• Conditional Mutation—wild type function except under
certain (permissive) conditions
– Ex. Temperature sensitive mutants show mutant phenotype only
at certain temperatures
• Leaky mutations— a missense amino acid change that
reduces but doesn’t eliminate protein function
 Mutagenesis

• The process of mutation

• Mutagen—anything that promotes or
causes mutations
– Chemical
– Physical
 Mutation-Causes
• Incorrect base pairing due to tautomeric shifts
• Removal of nitrogenous bases
• Alteration of nitrogenous bases
• Addition or deletion of nucleotides
• Single strand breaks
• Double strand breaks
• Crosslinking—covalent linkage between
bases
 Spontaneous Mutations
• Arise without mutagenic agents. DNA pol has
proofreading function, can remove
mismatched base
• Even if DNA pol misses a mismatch other
systems can recognize and repair it.
• Recognition?
– Hemimethylation-allows enzymes to distinguish
between parent and daughter strands.
 Tautomeric shifts
Nucleotides spontaneously under go a transient
rearrangement of bonding, e.g. a shift from
NH2 (amino form) to NH (imino form) or C=O
(keto) to C-OH (enol). Therefore, if any base in
a template strand exists in its rare tautomeric
form during DNA replication, misincorporation
in the daughter strand can result.
 Metabolite Mutagens
Chemicals that are metabolized to
electrophilic reagents: Aflatoxins,
benzo[a]pyrene
A mutagen is a physical or chemical agent
that causes mutations to occurs.
Mutagenesis is the process of producing a
mutation.
Mutant refers to an organism or a gene that
is different from the normal or wild type.
 Reversion and
the Ames test:

Reversion was used as a

means of detecting
mutagens and carcinogens-
the Ames test
 DNA Repair Mechanisms

(1) Repair by direct reversal: The simplest

mechanism. e.g. UV induced T-T dimer is
recognized by photolyase and is cleaved
into intact thymine (light dependent). This
is called photoactivation
 Excision Repair
(2) Excision Repair:
The most
ubiquitous repair
mechanism, which
can deal with a
large variety of
structural defects in
DNA.
 Recombinational Repair
(3) Recombinational repair (Postreplicational
repair): Occurs before excision repair has
happened or when excision repair can not
fix the problem
 The SOS response
(4) The SOS response: The SOS response system is only
active in response to some signal such as a blocked of
replication fork. In E. Coli, recA and lexA govern the
expression of a number of other genes involved in DNA
repair. This is an error-prone DNA repair mechanism and
result in higher than normal mutagenesis.
 SOS DNA Repair
1. DNA damage
2. RecA converted to RecA*
3. RecA* facilitated LexA self-cleavage
4. Increased synthesis of SOS proteins
5. Error prone repair induced
6. DNA damage repaired
7. RecA* returned to RecA
8. LexA no longer self-cleaved
9. LexA repressed SOS genes
10. LexA repress lexA gene expression
 Type of Mutations(I)
I. Point mutation:
A. Base substitution
Change in DNA
Transition: One purine replaced by a different
purine;or one pyrimidine replaced by a
diferent pyrimidine
A G T C
Transversion: A purine replaced by a pyrimidine
or vice versa
A T C G
 Type of Mutations (II)
Change in protein
1. Silent mutation: altered codon codes for the
same a.a. GAG (Glu) --->GAA (Glu)
2. Neutral mutation: altered codon codes for
functional similar a.a. GAG--->GAC (Asp)
3. Missense mutation: altered codon codes for
different dissimilar a.a. GAG ---> AAG (Lys)
4. Nonsense mutation: altered codon becsomes
a stop codon
GAG ---> UAG (stop)
 Type of Mutations (III)
B. Frameshift mutation: addition or deletion of one
base-pair result in a shift of reading frame and
alter amino acid sequence

1. Wild type: ATG ACC AGG TC

Met Thr Arg

2. Base addition: ATG ACA CAG GTC

Met Thr Gln Val
3. Base deletion: ATG ACA GGT C
Met Thr Gly
 Type of Mutations (IV)
II. Insertion

III. Deletion

IV. Translocation

V. Inversion