DNA Mutations,

Damage & Repair

Lecture 2

Recap
MCB2020F

Transcription and Translation

DNA Replication
(DNA makes a copy of itself)
Enzyme=
polymerase
DNA
Reverse transcription
Eg. retroviruses

Transcription of DNA to mRNA

Enzyme =
polymerase
RNA

Protein

Translation of mRNA to protein

Ribosomes = ribosomal RNA
proteins
mRNA template
tRNA brings in amino acids

Recap
MCB2020F

Transcription and Translation

in bacteria

= Ribosome binding site

NO INTRONS
Translated protein
AUG
Start codon

UAA
Stop codon

Recap
MCB2020F

Table of Standard Genetic Code

DNA Structure & Replication

COMPLEMENTARY BASE PAIRING

Adenine (Purine) base pairs with Thymine (Pyrimidine)
Guanine (Purine) base pairs with Cytosine (Pyrimidine)
DNA STRAND SEPARATION

NEW STRAND SYNTHESIS

Types of mutations

Macrolesions

Large disruptions to genome

Transposons

Duplications

Large deletions

Chromosomal inversions

Microlesions

Smaller changes (1-2 bases) to the genome

General Types of mutations of

bases

Base substitutions

Exchange one base for another base

Frameshift mutations

Insertion or deletion of bases

Types of mutations
1. Base substitutions
Purine (Pu)

Pyrimidine (Py)

Transitions
Pu replaces another Pu eg: A to G (or G to A)
Py replaces another Py eg: T to C (or C to T)

Transversions
Pu replaces a Py
Py replaces a Pu

eg: A to T/C or G to T/C

eg: T to A/G or C to A/G

Effects of Base substitution

(in coding region)
i)

Same sense mutation (silent mutation)

Because of degenerate genetic code

eg: serine = TCT (DNA)

UCU (mRNA)

TCC
UCC

T to C transition in DNA but same aa inserted

No physiological effect

Effects of base substitution

ii) Missense mutation
Base change leads to amino acid change
May affect protein function (or may not)
Depends on position in aa chain:
active site
co-factor binding site
protein folding
stability

If aa functionally conserved, maybe no change in

enzyme activity

Effects of base substitution

iii) Nonsense mutations
Generates stop codon
UAG or UAA or UGA (mRNA)
TAG or TAA or TGA (DNA)
Creates truncated protein
Very drastic consequences

May be less harmful if close to end of gene

Mutations Caused by Base

Substitutions
Silent
mutation

No mutation

Missense
mutation

Nonsense
mutation

TCA
AGT

TTA
AAT

TAA
ATT

TCG
AGC

UCA

UUA

UAA

UCG
Stop Codon

Ser

Leu

Ser

Effects of base substitution

in regulatory/recognition region

Lack of DNA-protein recognition or binding

Decreased/increased transcription

Inaccurate mRNA termination point

Inaccurate ribosome binding to mRNA

Inaccurate tRNA recognition of codons on mRNA

wrong aa incorporated

Types of mutations
2. Frameshift mutations
addition or deletion of bases

Effect depends on position and number of bases involved

WT DNA: ATG|CCA|TAC|TGG
WT mRNA: AUG|CCA|UAC|UGG
WT aa:
met pro tyr trp

Addition of 1 base?
mutant DNA: ATG|CCA|GTA|CTG|G
mutant mRNA: AUG|CCA|GUA|CUG|G
mutant aa:
met pro val leu and all other aas

Reading frame shifts

All subsequent aa change (or could cause stop codon)
Very drastic effects

How are mutations caused at the

DNA level?
Spontaneous errors

Tautomerism
Replication errors
Depurination
Insertion of DNA via MGE

Induced errors

Chemical
Chemicals
Pollutants
Physical
Radiation (UV, Gamma)

Tautomerism

Tautomerism is the relationship between two

structural isomers that are in chemical equilibrium
and readily change from one to another.

Nitrogenous base of template strand can takes on a

rare tautomeric form.

Bases typically exist in the keto form but can take

on either imino or enol form.

Tautomeric shifts lead to change in hydrogen

bonding characteristics of the bases.

Tautomerism

Tautomeric forms allow for transition mutations

to occur.

Tautomerism

Process requires two replication cycles - DNA

repair mechanisms will repair the anomalous base
pair formation in the first generation.

Depurination

Purines can become depurinated (lose their base)

to form a apurinic site.

Apurinic site does not base pair normally and

repair enzymes may randomly add the wrong
base.

Can result in either a transition or transversion

after replication.

Depurination