MUTATION

 KINDS
 MECHANISMS
 AGENTS
 REPAIR MECHANISMS
 Mutation
（突变）
Replication Mutagenesis
Fidelity
( 复制的忠实
Mutagens
性）
（诱变剂）
 Mutation

Concept: Permanent, heritable alterations

in the base sequence of DNA

Arise either through spontaneous errors in DNA

replication or meiotic recombination or as a
consequence of the damaging effects of physical or
chemical agents on the DNA.
 Mutation
Point mutation: is the simplest mutation-a single base
change.

1, Transition ( 转换 ): Purine or pyrimidine is replaced

by the other AG T C

2, Transversion ( 颠换 ): a purine is replaced by a

pyrimidine or vice verse
A T or C T  A or G
G T or C C  A or G
 Mutation
Effects of a point mutation
The sites Types Phenotypic
effects
•Noncoding DNA
•Nonregulatory DNA Silent mutation No
•3rd position of a codon

Missense Yes or No
Coding DNA  altered AA
mutation

Coding DNA  stop codon

 truncated protein Nonsense mutation Yes
 Mutation
Insertions or deletions

The addition or loss of one or more bases in a DNA region

Frameshift mutations
The ORF of a protein encoded gene is changed so that the C-
terminal side of the mutation is completely changed.
Consequences of base
substitutions

 Silent mutation: base change results in no

change of the amino acid sequence of the
translated protein
 Silent mutations have no effect on phenotype
 A result of the fact that multiple codons can
code for the same amino acid
 E.g., AGU and AGC both code for Serine
Consequences of base
substitutions
 Missense mutation: base change results in the
change of an amino acid in the translated protein
Consequences of missense
mutations
 The amino acid substitution induced by the
missense mutation may have no effect on
the function of the protein OR
 It may abolish the activity of the protein or
alter its function having an effect on
phenotype
Example: sickle cell disease in humans is due
to a missense mutation in the gene for globin.
As a result the shape of red blood cells is
altered affecting their movement through
capillaries.
Consequences of base
substitutions
 Nonsense mutation: base change generates a
stop codon in place of that coding for an amino
acid
 Results in production of a truncated protein.
Usually results in a non-functional protein
Consequences of frameshift
mutations
 Frameshift mutation: addition or deletion of one or
more bases
 Results in misreading of the codons (changed
reading frame)
 Almost always results in long stretches of altered
amino acids and the production of inactive protein
Causes of mutations

 Spontaneous mutations
 Occur in the absence of mutation causing
agents
 Due to occasional mistakes in DNA replication
 Induced mutations
 Caused by mutagens, agents such as
chemicals and radiation which induce
mutations
Chemical mutagens
 Example: Nitrous acid alters adenine such that it
pairs with cytosine instead of thymine
 Replication fidelity
The need to preserve the meaning of the genetic
information from one generation to the next, so the error
rate of DNA replication is much lower. E.g. Spontaneous
errors in DNA replication is very rare, one error per 1010
base in E. coli.
Molecular mechanisms of the replication fidelity

1. DNA polymerase: Watson-Crick base pairing

2. The occasional error can be detected and repaired by 3’
5’ proofreading exonuclease.
3. RNA priming: proofreading the 5’ end of the lagging strand
4. Mismatch repair mechanism(F3)
 Proofreading
by
E. coli polymerase
(Figure from Gene VII)
 Mutagens
The physical and chemical agents of causing DNA damage that
can be converted to mutations.
1, Physical mutagens
High-energy ionizing radiation: x-rays and -rays  strand breaks and base/sugar
destruction (cause extensive chemical alterations)
Nonionizing radiation: UV light pyrimidine dimers from adjacent

2, Chemical mutagens
Base analogs: direct mutagenesis; Nitrous acid: deaminates C to produce U and
guanine analog; Alkylating agents and Arylating agents produce lesions (indirect
mutagenesis); Intercalators: insertion and deletion mutations
 Base analogs: derivatives of the normal bases with
altered base pairing properties.

Nitrous acid: deaminates C to produce U, resulting in G·C

 A·U
Chemical mutagens
 Example: Nitrous acid alters adenine such that it
pairs with cytosine instead of thymine
Chemical mutagens
 Example: Ethidium bromide inserts
between bases causing frameshift mutation

 Example: Nucleotide analogues-substitute

for a base but have different pairing
properties. Some can be used as anti-viral
or tumour therapy
Radiation
 Ionizing radiation e.g., Xrays and gamma rays
Causes the formation of ions that can react with
nucleotides (causing base changes) and the
deoxyribose-phosphate backbone (causes
chromosomes to break).

 UV radiation
Induces formation of
covalent bonds between
adjacent thymines to form
thymine dimers which can not be replicated
 Mutagenesis
molecular process in which
the mutation is generated,
including direct and
indirect
 E.g. AGCTTCCTA
TCGAAGGAT
1. Base analog
1, Direct OH incorporation
mutagenesis H Br
The stable, :G AGCTBCCTA
unrepaired O TCGAAGGAT
base with 2. 1st round
altered base enol form of replication
pairing
properties in AGCTTCCTA AGCTBCCTA
the DNA is TCGAAGGAT TCGAGGGAT
fixed to a
H Br
mutation : A3. 2nd round
during DNA O of replication
replication.
Keto form AGCTBCCTA AGCTCCCTA
TCGAAGGAT TCGAGGGAT
5-BrU
A·TG·C transition
2, Indirect mutagenesis
The mutation is introduced as a result of an error-
prone repair.
Translesion DNA synthesis to maintain the DNA
integrity but not the sequence accuracy: when
damage occurs immediately ahead of an advancing
fork, which is unsuitable for recombination repair
(F4), the daughter strand is synthesized regardless of
the the base identity of the damaged sites of the
parental DNA.
E 。 g 。 E. coli translesion replication: SOS
response
 DNA damage and repair

Physical and chemical Mutagens

minor or DNA damage The lesions of extensive,

moderate lesions (lesions) right ahead of
Replication Fork
Error-free Error-prone
Repairing repairing

Completely repaired mutations

 DNA damage
DNA lesion is an alteration to the normal chemical or
physical structure of the DNA.
1, Oxidative damage DNA lesions 2, Bulky adducts
( 氧化损伤） (DNA 损害） ( 加合物）

1. Occurs under
Normal condition UV light
2. Increased by (physical mutagens)
3 ， Alkylation Carcinogen
ionizing radiation ( 烷基化作用）
(physical mutagens) (Chemical mutagens)
Alkylating agents
(Chemical mutagens)
 F2 DNA damage-2
The biological effect of the unrepaired DNA lesions

Physical distortion in The altered chemistry of the bases

the DNA may lead to loss of base pairing or
altered base pairing. If such a
lesion was allowed to remain in
Blocks replication the DNA,
And/or transcription Living cell

Lethal A mutation could become fixed

(cell death) by direct or indirect mutagenesis

Mutagenic
 F2 DNA damage-3
Spontaneous DNA lesions
Because of inherent chemical reactivity of the DNA and
the presence of normal, reactive chemical species
within the cell

1. Deamination ( 转氨作用） : CU; methylcytosine T,

hard to be detected
2. Depurination ( 脱瞟呤作用 ) : break of the glycosylic bond,
non-coding lesion.
3. Depyrimidine ( 脱嘧啶作用 )
 F2 DNA damage-4
Oxidative damage
1, Is a kind of DNA lesions caused by reactive oxygen species
such as superoxide and hydroxyl radicals, which occurs under
normal conditions.
2, The level of this damage can be INCREEASED by hydroxyl
radicals from the radiolysis of water caused by ionizing
radiation.

Oxidation products
 F2 DNA damage-5
Alkylation

Alkylating agents are electrophilic chemicals which

readily add alkyl (e.g. methyl) groups to various positions
on nucleic acids distinct from those methylated by
normal methylating enzymes such as methylmethane
sulfonate （甲基甲烷磺酸盐） and ethylnitrosourea ( 乙
基亚硝基脲 ).
alkylating agents

Alkylated bases
 F2 DNA damage-6
Bulky adducts

A kind of DNA lesions that distort the double

helix and cause localized denaturation, for
example pyrimidine dimers and arylating agents
adducts. These lesions disrupt the normal
function of the DNA.
Cyclobutane pyrimidine dimer( 嘧啶二聚体 )

Guanine adduct of benzo[a]pyrene

Aromatic
arylating agents

Covalent adducts
 F3 DNA repair-1
Photoreactivation

Cyclobutane pyrimidine dimers can be monomerized again

by DNA photolyases (photoreactivating enzymes) in the
presence of visible light.

Direct reversal of a lesion and is error-free

 DNA repair
 Prokaryotic and Eukaryotic
cells have the capacity to
repair DNA
 DNA photolyase recognizes
and repairs thymine dimers
using energy from light (light
repair, photoreactivation)
 A general excision repair
system excises incorrect
bases leaving a gap for DNA
polymerase I and ligase to fill
 F3 DNA repair-2
Alkyltransferase
Removes the alkyl group from mutagenic O6-
alkylguanine which can base-pair with T. The alkyl
group is transferred to the protein itself and
inactivate it.

Direct reversal of a lesion and is error-free

 F3 DNA repair-3
Excision repair

1. There are two forms, nucleotide excision

repair (NER) and base excision repair
(BER).
2. Is a ubiquitous mechanism repairing a
variety of lesions.
3. Error-free repair
Nucleotide excision repair

1. An endonuclease cleaves
DNA a precise number
of bases on either sides
of the lesions (in E.coli
UvrABC endonulcease
removes pyrimidine dimers)
2. Excised lesion-DNA
fragment is removed
3. The gap is filled by
DNA polymerase I and
sealed by ligase
Base excision repair
1. modified bases are recognized
by relatively specific DNA
glycosylases which cleave the
N-glycosylic bond between the
altered base and the sugar),
leaving an apurinic or
apyrimidinic (AP) site.
2. An AP endonuclease then
cleaves the DNA at this site
and a gap may be created by
further exonuclease activity.
The gap is generally larger in
NER and can be as small as
one nucleotide in BER.
3. The gap is filled by DNA
polymerase I and sealed by
ligase
 F3 DNA repair-4
Mismatch repair

Is a specialized form of excision repair that

deals with any base mispairs produced
during replication and that have escaped
proofreading.
error-free
 The parental strand is methylated at N6
position of all As in GATC sites, but
methylation of the daughter strand lag a few
minutes after replication

MutH/MutS recognize the mismatched

base pair and the nearby GATC

DNA helicase II, SSB, exonuclease I

remove the DNA fragment including the
mismatch

DNA polymerase III & DNA ligase

fill in the gap

Expensive to keep the accuracy

Molecular Biology

DNA Repair Mechanisms

The proteins of DNA replication also play roles in the life-preserving repair
mechanisms, helping to ensure the extract replication of template DNA.
alkylating agents

Alkylated bases
 F2 DNA damage-6
Bulky adducts

A kind of DNA lesions that distort the double

helix and cause localized denaturation, for
example pyrimidine dimers and arylating agents
adducts. These lesions disrupt the normal
function of the DNA.
 F2 DNA damage-6
Bulky adducts

A kind of DNA lesions that distort the double

helix and cause localized denaturation, for
example pyrimidine dimers and arylating agents
adducts. These lesions disrupt the normal
function of the DNA.