Mutations

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• Mutation is change in DNA genetic makeup
• Mutagen is an agent of substance that can bring
about a permanent alteration to the physical
composition of a DNA gene such that the
genetic message is changed
• Mutations were initially characterized as altered
phenotypes
• But now they are understood at molecular level

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• Small changes in DNA are called Microlesions
• Smallest microlesion is called point mutation
• Point mutations affect only one base pair in a
given location

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• Larger mutations are called Macrolesions
• Macrolesions include
– Insertions
– Deletions
– Inversions
– Duplications
– Translocations

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Types of mutations

• Mutations occur in two ways

1. Spontaneous mutations
2. Induced mutations

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1. Spontaneous Mutations

• These mutations arise without exposure to

external agents
• These mutations may result from errors in DNA
replication or from mobile genetic elements such
as transposons

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Tautomerism
• Replication errors occur when nitrogenous base
of template nucleotide takes on a rare
tautomeric form
• Tautomerism is a relationship between two
structural isomers that are in chemical
equilibrium and readily change into one another

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• Bases typically exist in keto form
• However they can takes imino or enol form
• These tautomeric shifts change hydrogen
bonding characteristics of bases which lead
to stable alteration of nucleotide sequence
• These changes are known as transition
mutations

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• In transition mutations, purine for purine and
pyrmidine for pyrimidine substitution
• In transversion mutations, purine is substituted
for pyrimidine and pyrimidine for purine
• Transversion mutations are rare
• Sometimes, addition or deletion of nucleotides
occur
• These generally occur where there is a short
stretch of same nucleotide

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Depurination
• Loss of a purine base (A or G) to form an
apurinic site (AP site)
• The formation of an apurinic site does not base
pair normally and may cause a transition type
mutation after next round of replication
• Similarly, pyrimidines can be lost forming an
apyrimidinic site

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• Sometimes reactive forms of oxygen such as
free radicals and peroxides produced during
aerobic metabolism
• These may alter DNA bases and cause
mutations
– e.g. guanine can be converted to 8-oxo-7,8-
dihydrodeoxyguanine, which often pairs with adenine
rather than cytosine during replication

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• Finally, spontaneous mutations can result from
insertion of DNA segments into genes
• Insertions usually inactivate genes
• They are caused by movement of insertion
sequences and transposons

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2. Induced mutations

• Mutagens can be classified according to their

mode of action
• Chemical mutagens
– Base analogs, DNA modifying agents, intercalating
agents
• Physical mutagens
– Radiations (UV, X-rays, gamma)

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Base analogs
• They are structurally similar to normal
nitrogenous bases and can be incorporated into
growing polynucleotide chain during replication
• Once in place, these compounds typically exhibit
base pairing properties different from bases they
replace and can eventually cause a stable
mutation
• e.g. 5-bromouracil (5-BU), an analog of thymine
• It undergoes a tautomeric shift from normal keto
to an enol form

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DNA-modifying agents
• These agents change a base’s structure and
therefore alter its base pairing characteristics
• Some mutagens react with selective bases and
produce a specific kind of DNA damage
– e.g. methyl-nitrosoguanidine (an alkylating agent that
adds methyl groups to guanine, causing it to mispair
with thymine
• A subsequent round of replication could then result in a
GC-AT transition
– Hydroxylamine
• It hydroxylates the C4 nitrogen of cytosine, causing it to
base pair like thymine

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Intercalating agents
• These agents distort DNA to induce single
nucleotide pair insertions and deletions
• These mutagens intercalate between bases
resulting in a mutation possibly through
formation of a loop in DNA
• e.g. acridine orange, ethidium bromide

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• Many mutagens (including carcinogens) directly
damage bases so severely that hydrogen
bonding between base pairs is impaired or
prevented, and damaged DNA can no longer act
as template for replication
– e.g. UV generates thymine dimers between adjacent pyrimidines
• Ionizing radiations and carcinogens such as
fungal toxins aflatoxin B1 and other
benzo(a)pyrene derivatives

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Effects of mutations

• Forward mutation: Change from wild type to

mutant form is called forward mutation
• Reverse mutation: A forward mutation can be
reversed by a second mutation that restores the
wild type phenotype. When the second mutation
is at the same site as the original mutation
• A true reversion converts the mutant nucleotide
sequence back to wild type sequence. If the
second mutation is at a different site than the
original mutation, it is called Supressor mutation

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• Supressor mutation may be within same gene
called as intragenic suppressor mutation

• Supressor mutation may be in different gene

called as extragenic suppressor mutation

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Mutations in protein coding gene

• Point mutations in protein coding genes can

affect protein structure in a variety of ways
• Point mutations are named according to if and
how they change the encoded protein
• The common type of point mutations are as
follow
– Silent mutation
– Missense mutations
– Non sense mutations
– Frameshift mutations

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• Conservative results in a new amino acid that has similar properties
as the original.
• Non-conservative results in a new amino acid with very different
properties than the original.
• Conservative missense mutations are also known as neutral
missense mutations
Macrolesions

• Insertions
• Deletions
• Inversions
• Duplications
• Translocations

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Insertion

• It is the addition of one or

more nucleotide base pairs into
DNA sequence
• Insertions can be anywhere in
size from one base pair
incorrectly inserted into a DNA
sequence to a section of one
chromosome inserted into
another

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• On a chromosomal
level, an insertion
refers to insertion of a
larger sequence into
a chromosome
• This can happen due
to unequal crossing
over during meiosis

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Deletion

• Deletion is the loss of genetic material

• Any number of nucleotides can be
deleted, from a single base to an entire
piece of chromosome
• Sign: Δ
• Deletions can be caused by errors
in chromosomal crossover during
meiosis
• This causes several serious genetic
diseases
• Deletion also causes frameshift
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Inversion

• This mutation causes

a reversal in the order
of a segment of a
chromosome within
the chromosome, or a
gene

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Duplications

• A major mechanism through which

new genetic material is generated
during molecular evolution
• It can be defined as any duplication
of a region of DNA that contains
a gene, it may occur as an error
in homologous recombination

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Translocation

• Abnormality caused by
rearrangement of parts
between non homologous
chromosomes
• There are two main types
– Reciprocal (also known as non-
Robertsonian)
– Robertsonian

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The End

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