RECOMBINANT DNA202
DNA MUTATION
Dr Danisile Tembe, SLS Room 05-105, Email: [email protected]
WHAT IS DNA MUTATIONS?
➢Mutation is a change in the nucleotide sequence of an organism’s DNA
• These changes are inheritable
➢The change of a single nucleotide in the DNA template strand results to
• an altered mRNA & the production of abnormal protein
➢Mutations of one or few base pairs usually results from errors during replication
or damaged nucleotides
• Such mutations include: point mutations, small insertions and/or deletions
WHAT IS DNA MUTATIONS?
➢Mutations in more base pairs/ large section of DNA sequence are due to chromosomal
rearrangement that arise due to errant recombination
➢Such mutations include: duplication mutation, inversion mutations, and translocation
mutations
TYPES OF DNA MUTATIONS DURING REPLICATION
1. Point mutation/substitution
➢Change in a single base pair
• Caused when incorrect nucleotide is incorporated by a DNA polymerase
• It falls into two categories: transition mutation and transversion mutation
5’AGCTA3’
3’TCGAT5’
exchange of a purine-
Transitions Transversions
pyrimidine base pair for the replacement of a purine-pyrimidine base
other purine-pyrimidine base 5’AGTTA3’ 5’AGGTA’ pair wit pyrimidine- purine base pair for
pair C-G become T-A 3’TCAAT5’ 3’TCCAT5’ example C=G becomes either G=C or A=T
OR
5’AGATA’
3’TCTAT5’
TYPES OF DNA MUTATIONS DURING REPLICATION
1. Point mutation/substitution
➢Point mutation in the protein coding region of a gene can result in an altered protein with
partial or complete loss of function
➢Point mutation in a protein coding region can be classified by their effect on the protein
sequence
➢We will focus on small scale mutations occurring within a gene
➢Small scale mutation can be divided into two:
➢ Single nucleotide - pair substitution & Nucleotide-pair insertion or deletion
(1) NUCLEOTIDE SUBSTITUTION
➢ Nucleotide-pair substitution is the replacement of one nucleotide & its partner with another
pair of nucleotides
➢ This mutation can be divided into 3:
Silent mutation: has no observable effect on the phenotype
• E.g. a mutation that results in a codon that codes for the same amino acid
(1) NUCLEOTIDE SUBSTITUTION
➢ Missense mutation: mutation/substitution that changes one amino acid to another
• Results in a codon that codes for a different amino acid
• Might have little effects on the protein
(1) NUCLEOTIDE SUBSTITUTION
➢ Nonsense mutation: mutation that changes amino acid codon to a stop codon
• Causes translation to be terminated prematurely
• Resulting in a shorter & usually non-functional protein
(2) NUCLEOTIDE INSERTIONS & DELETIONS
➢ Insertion and deletions: additions and losses of nucleotide pairs in a gene
• These mutations have major effects on the resulting protein more often than a
substitutions
➢ May change the reading frame of the genetic message
• This mutation is known as frameshift
➢ Frameshift occurs when the number of nucleotides added or deleted is not in a multiple of 3
• All nucleotides downstream to the insertion or deletion will be improper grouped into
codons
• This result to extensive missense mutation, usually resulting to nonsense mutation that
lead to premature termination
(2) NUCLEOTIDE INSERTIONS & DELETIONS
ALTERATIONS THAT LEAD TO DNA MUTATIONS
➢Spontaneous mutations: occurs without known cause
➢Induced mutations: occurs due to exposure to physical or chemical agents
• Mutagens – chemicals and radiation
SPONTANEOUS MUTATIONS
1. DNA Tautomers:
➢occurs when the base takes on either an imino or enol form instead of the normal keto form
2. Trinucleotide repeats:
➢ a region of chromosome that is highly repeated
• bases are highly repeated in a DNA sequence
SPONTANEOUS MUTATIONS
3. Depurination
➢ The loss of a purine base by spontaneous hydrolysis of the glycosidic bond
4. Deamination
➢ The removal of an amino group(-NH2) from a compound
• All molecules that contain amino group are possible targets for hydrolytic attack
• 3 nucleotide bases contain an amino group (cytosine, adenine, and guanine) and they can
deaminated by hydrolysis
• Deamination of cytosine is the common one and it results in uracil
➢Spontaneous hydrolysis reactions caused by random interactions between DNA and the
water molecules around it
DEPURINATION AND DEAMINATION
INDUCED MUTATIONS: CHEMICAL MUTAGENS
➢Induced mutations are caused by
chemical mutagens or radiation
1. Base analogues:
• They resemble the nitrogenous bases
in structure and are incorporated in
DNA
• Example is 5-bromodeoxyuridine (BrdU)
INDUCED MUTATIONS: CHEMICAL MUTAGENS
2. Base modifying-agents:
• react with with a nitrogenous base and alter their structure
• Base modifying agents normally results in depurination.
3. Intercalating agents:
• alters the shape of the DNA double helix, leading to small nicks in the DNA
INDUCED MUTATIONS: RADIATION
• Change in DNA triggered the by UV light and they cause pyrimidine dimmer formation
• For example in this case we have Thymine binding with thymine
