D.

Types of mutation
1. Point mutation.
Affecting non coding regions: e.g. Promoter/operator e.g. -10 TATAAT sequence in promoter Affecting CODING SEQUENCE or Open Reading Frame (ORF) sequence. AAT DNA UUA mRNA Leu amino acid

Can mutate to..

CUA Leu

GUA Val

AUA Ile UGA Stop

UCA Ser UAA Stop

UUC Phe

UUG Leu

UUU Phe

UCA Ser

M J Larkin Biology & Biochemistry. The Queens University of Belfast.

D. Types of mutation. Cont.

STOP leads to truncated protein and termination of translation (Note: Transcription termination involves inverted repeats and role of rho protein factor) Some codons are NONSENSE (i.e. normally no tRNA) UAG Amber (after discoverer Bernstein :German for Amber) UAA Ochre UGA Opal

2. Frameshifts. Addition or deletion of bases leading to

altered sequence beyond the sequence change.
M J Larkin Biology & Biochemistry. The Queens University of Belfast.

D. Types of mutation. Cont.

3. Deletions
Arise spontaneously where there are direct repeats of over 45 base pairs. More frequent when there is more sequence repeated (see later excision of Transposons and IS elements)

i.e.
3 5

Replication slippage 5 3

Gap maybe up to a few 1000 bps

Recombination and deletion

M J Larkin Biology & Biochemistry. The Queens University of Belfast.

E. Reversion and Suppression

Back mutation to original sequence is rare Suppressor mutations relieving mutations within coding regions more common Leads to insertion of amino acid at stop codon
e.g. CAG Gln UAG STOP Amber AUC Protein may or may or be functional Mutation in another tRNA leads to incorporation of a different amino acid

GUC
Gln tRNA

M J Larkin Biology & Biochemistry. The Queens University of Belfast.

F. Mechanisms of mutation
Spontaneous mutation. ERRORS IN REPLICATION Usually only AT GC pairing allowed Repair systems are present How do mis-matches or mis-pairings happen ? Alternative Tautomeric forms of A and T occur
(In evolution we are all prisoners of simple chemistry)

Dale Ch 3
M J Larkin Biology & Biochemistry. The Queens University of Belfast.

F. Mechanisms of mutation. Cont.

2 x H Bonds
3 x H Bonds
O H
N

H O
N

CH3 N H

CH3 N H H

Thymine in its KETO form Will bind to Adenine TA pairing as expected during replication

Thymine in its ENOL form. Binds Guianine. TG pairing during replication

M J Larkin Biology & Biochemistry. The Queens University of Belfast.

F. Mechanisms of mutation. Cont.

SIMILARLY
NH2
N

Still 2 x H Bonds 2 x H Bonds

NH
N

Adenine in its AMINO form Will bind to Thymine. TA pairing as expected.

Adenine in its IMINO form. Binds Cytosine. AC pairing.

Forms in equilibrium with about 1 in 104 or 105 molecules in the ENOL or IMINO forms. Therefore natural transition per base per generation is also about 1 in 104 to 105.
M J Larkin Biology & Biochemistry. The Queens University of Belfast.

F. Mechanisms of mutation. Cont.

Must also have segregation after replication

1st replication
A T G T

2nd replication

G C
A T

This would happen about 1 in 104 to 105 ! The potential for many deleterious mutations is high i.e. GENETIC LOAD is too high
Dale Ch 3
M J Larkin Biology & Biochemistry. The Queens University of Belfast.

F. Mechanisms of mutation. Cont.

WORKING AT ANOTHER LEVEL AFTER REPLICATION IS Mismatch repair. A form of excision repair (see later). DNA adenine methylation is involved in recognising sections of DNA to be repaired after replication.

DNA can also become damaged due to a variety of influences. Chemical mutagens Ionising radiation (such as X rays, rays and UV254nm) Repair of this damage rapidly can lead to mutagenic effects.
Dale Ch 3
M J Larkin Biology & Biochemistry. The Queens University of Belfast.

F. Mechanisms of mutation. Cont.

Chemical mutagens Base analogues: substitute for normal base but are not proofread - mispairing 2-aminopurine: Adenine analogue which pairs with cytosine 5-bromouracil: thymine analogue pairs with guanine Modification of bases: Do not require replication and induce DNA repair. Nitrous acid (deaminates); hydroxylamine (reacts with cytosine) Alkalyating agents transfer alkyl groups to bases. Very powerful; ethyl methane sulphonate (EMS); N-methyl, Nnitrosoguanidine (NTG) Intercalating agents: additions or deletions caused acridine orange; ethidium bromide

M J Larkin Biology & Biochemistry. The Queens University of Belfast.

F. Mechanisms of mutation. Cont.

Proofreading carried out by the 3 to 5 exonuclease activity of the DNA Polymerases III and I.
Removal of mismatch 3 - 5 digestion by polymerase

AG or CT Mismatch

Polymerisation in 5 - 3 direction
Polymerase

THIS WORKS AT ONE LEVEL

M J Larkin Biology & Biochemistry. The Queens University of Belfast.