Mutation

Introductory article
Article Contents

Raymond Devoret, Curie Institute, Paris, France

. Introduction

A mutation is a DNA change that is recorded durably and passed on to the offspring. A
number of mutations in germ cells have detrimental effects for they often cause hereditary
diseases. Likewise, mutations that accumulate in body cells can generate cancers.
Nevertheless, a large number of mutations are neutral in their normal environment. The
few advantageous mutations that occur are generally maintained. Mutations that arise at
random are constantly sifted through by a selective environment. This dual process works
as the driving force of evolution.

. Nature of Mutations
. Most Mutagens are DNA-damaging Agents
. DNA Lesions and Mutations
. Repair of DNA Lesions
. Mutation Types, Reversions and Location
. Synonymous Mutations are Neutral
. Mechanisms Generating Mutations
. A Majority of Mutagens are Carcinogens. The Ames
Test

Introduction

. Carcinogens Leave No Definite Mutational Signatures

Deoxyribonucleic acid (DNA) is the essential component

of chromosomes in all known cells. Each chromosome is
made of an extremely long DNA molecule with two intertwined complementary strands (a double helix). Each
strand consists of a poly (phosphatesugar) backbone,
from which project adenine (A), cytosine (C), guanine (G),
and thymine (T) bases at each sugar. The two strands form
a duplex maintained by hydrogen bonds between complementary bases: A binds to T and G to C. The linear succession of combinations of A, C, G and T bases constitutes
a genetic sequence (Figure 1).
The DNA of the whole set of chromosomes in a cell is
called the genome. A bacterium like Escherichia coli, a
natural resident of the intestinal tract of mammals, has one
chromosome, whereas human cells carry 23 chromosome
pairs. In general, the cells of complex organisms like mammals have more chromosomes per cell than lower organisms like microbes.
The main property of DNA is to code for the hereditary
traits that are passed from parents to progeny. Recently,
the long string of bases that constitutes the genome of
many organisms, including humans, has been identied.

doi: 10.1038/npg.els.0001882

This achievement has established the precise chromosome

positioning of many genes that control hereditary traits. It
was discovered that the genes that code for proteins performing DNA replication are similar in all living organisms. This emphasizes the strong conservation of the
primary mechanism of cell division. Furthermore, the
genes that control mutagenesis (the process leading to the
production of mutations) are very much alike in different
organisms. DNA is able to mutate so that living organisms
can adapt to new environments.

Nature of Mutations
When a mutation occurs within a gene, the protein encoded
by the gene is often altered (Figures 1b,c and 2c); this alteration may produce a visible change in the displayed char-

(a)

AAGGCAAACCTACTGGTCTTATGT original sequence

(b)

AAGGCAAATCTACTGGTCTTATGT transition

(c)

AAGGCAAACCTACTGCTCTTATGT transversion

(d)

Del(ACCTA)
AAGGCAAvCTGGTCTTATGT deletion of ACCTA

(e)

Ins(AAAGC)
AAGGCAAACCTACTvAAAGCvGGTCTTATGT insertion of AAAGC

Figure 1 Types of mutation. (a) Wild-type original sequence; (b) transition from C to T; (c) transversion from G to C; (d) deletion (Del) of the sequence
ACCTA, the _ sign indicates from where it has been removed; (e) insertion (Ins) of the sequence AAAGC, the two _ signs indicate where the sequence has
been inserted.
ENCYCLOPEDIA OF LIFE SCIENCES 2004, John Wiley & Sons, Ltd. www.els.net

Mutation

acteristics (phenotype) of the organism studied. The actual

mutation itself (genotype) is invisible to the naked eye.
Since a mutation is a DNA change, it must be dened in
relation to a preceding, parental DNA sequence (Figure 1a).
Note, however, that the parental genome sequence may
already carry mutations; hence, geneticists have agreed to
dene a standard genetic sequence for each organism studied.
Three broad classes of mutation have been dened to
characterize their effects on the organisms in which they
arise: deleterious, neutral and advantageous. These qualications are valid only in relation to a specific environment. For example, a human newborn has a 1/1600
probability of suffering from cystic brosis, a disease that
prevents, in particular, the uidity of lung secretions. This
occurs if the child has received the two mutated cystic brosis gene copies from its parents. It means that 1/40
people carries one copy of the mutated gene; the aficted
individual has a normal phenotype but exhibits resistance
to acute infectious diarrhoea. The high frequency of that
mutation in the population can be explained by its selective
advantage. People die of cholera because a 20-litre diarrhoea per day kills them. People having one mutated allele
of the cystic brosis gene survive because they lose much
less water when infected with the cholera germ. In countries
where cholera has long been endemic, one mutated cystic
brosis allele can protect an individual against death from
cholera.

Most Mutagens are DNA-damaging

Agents
Naturally occurring mutations arise at low frequency. To
obtain more mutations the geneticist Herman Muller irradiated germ cells of Drosophila with X-rays and discovered
that radiation increases the frequency of mutations. Soon,
it was found that chemicals such as mustard gas (yperite)
also produce mutations at high frequency. Radiations and
some chemicals produce mutations: they are called mutagens. Both X-rays and yperite break chromosomes. This
property has been exploited in radiation therapy and
chemotherapy for destroying tumours. Likewise, ultraviolet light and many chemicals alter DNA in numerous
ways; these alterations are called DNA lesions.

DNA Lesions and Mutations

How can DNA-damaging agents produce mutations? In
other words, how can a DNA lesion give rise to a mutation?
DNA lesions display more than a hundred types of forms
and shapes, yet 85% of the resulting mutations are just a
2

change in one of the four known DNA bases; for example,

adenine may change into cytosine, guanine or thymine.
Most chemicals, for example benzopyrene found in tobacco smoke, when absorbed are metabolized into compounds that stick to DNA to form adducts (grafts). DNA
lesions are depicted with the vocabulary of chemistry,
whereas mutations are described with a simple alphabet of
four characters A, T, C and G, constituting the primary
code of life.
It is wrong to consider DNA lesions as mutations. A
whole set of biochemical reactions must proceed on the
damaged DNA to restore a proper DNA sequence. Note
that mutations are only observed in cells that have survived
DNA damage: dead cells carry numerous lesions that have
failed to be repaired.

Repair of DNA Lesions

Most DNA lesions block DNA replication. For a cell to
survive, DNA lesions must either be removed from the
chromosomes by repair processes or be bypassed by specific DNA polymerases. The polymerases generate a proper complementary single strand that will serve as the
template to restore a double helix in a second round of
replication.
Living organisms use at least 12 main repair processes to
remove DNA lesions or to bypass them. Repair enzymes
work with optimal efciency when the number of lesions is
low.
Interestingly, as soon as lesions are produced on DNA,
they provoke the SOS response, which induces a cascade of
repair processes. As repair fails at some DNA lesions, a
mutagenic DNA polymerase inserts a base across the lesion, even if its coding value is incorrect. Structural restoration of a DNA double helix is primordial to permit
replication and cell division.

Mutation Types, Reversions and

Location
Since DNA is universal, one nds the same alphabet of four
bases forming the genetic apparatus of all species. Mutation types are similar wherever they arise.

Point mutations
The most frequent are base substitutions. Substitutions between purines, adenine (A) and guanine (G), or between
pyrimidines, cytosine (C) and thymine (T) (Figure 1b) are
called transitions. An AT base pair may give rise to a GC
(AT ! GC) and, conversely, a GC base pair to an AT (GC

Mutation

! AT); the two-way reactions are represented as AT $

GC.
Transversions are substitutions of a purine for a pyrimidine and vice versa (Figure 1c). There are four reciprocal
base pair changes: AT $ CG, AT $ TA, GC $ TA and
GC $ CG. Base substitutions occurring in protein-coding
regions affect the expressed protein except when the change
is in the third base of a codon (see neutral or synonymous
mutation, Figure 2a).
A mutation in a gene that may not cause any amino acid
change in the expressed protein is called silent or synonymous, as it does not induce any change in the coded protein.
In Figure 2a, the valine amino acid is conserved in the protein sequence, while molecular techniques reveal a DNA
change.
A nonsynonymous mutation alters an amino acid, resulting in a missense or a nonsense mutation. A missense
mutation modies the affected codon, specifying an amino
acid different from the one previously encoded, for instance, valine (Val) becomes phenylalanine (Phe) (Figure 2b). A nonsense mutation changes a codon into one
of the three termination codons TAG, TAA or TGA.
Then, a truncated protein is produced that rarely has activity, for instance, lysine (Lys) becomes a stop condon
(Figure 2c).

Deletions and insertions

Examples of deletions and insertions are seen in Figure 1d,e.
Moreover, an unequal crossing-over between two chromosomes results in the deletion of a DNA segment in one
chromosome and an addition in the other.

Frameshifts
A small deletion or insertion within a gene, each involving a
number of bases which is not a multiple of three, causes a
shift in the reading frame, denoted frameshift (Figure 3a). In
this case, the coding sequence downstream is read in a
wrong phase. This new phasing changes the encoded
amino acids or else a new stop codon may be brought upstream into phase, thus producing a shortened protein.
Figure 3a shows a protein that may not have normal activity. Figure 3b also shows that addition of a base produces a
+1 frameshift, removing a preexisting stop signal and giving rise to an elongated protein.

DNA rearrangements
Deletions of segments of genes or sets of genes reduce or
eliminate protein functions (Figure 3). In addition, insertions of large DNA segments can also occur, as can duplications, inversions and other more complex DNA
rearrangements. All these processes are the results of recombination between DNA sequences sharing some similarity. Agents that break chromosomes, such as X-rays
and yperite, facilitate DNA rearrangements.
For instance, when a piece of human chromosome 14
carrying the Bcl2 gene, which prevents cell death, is translocated to chromosome 18, then the Bcl2 gene becomes
expressed continuously. Cells will consequently produce
mutated immunoglobulins, which are normally eliminated
from the body by cell death but kept alive here because of
the Bcl2 translocation. Thus, a pool of unhealthy cells
builds gradually; point mutations accumulate in those unhealthy cells, which will eventually give rise to cancer cells.

(a) Ile
ATA

Cys
TGT

Ile
ATA

Lys
AAG

Ala
GCA

Leu
CTG

Val
GTC

Leu
CTG

Leu
TTA

Thr amino acid sequence

ACA DNA sequence

ATA
Ile

TGT
Cys

ATA
Ile

AAG
Lys

GCA
Ala

CTG
Leu

GTA
Val

CTG
Leu

TTA
Leu

ACA base substitution

Thr synonymous amino acid

(b) Ile
ATA
ATA
Ile

Cys
TGT
TGT
Cys

Ile
ATA
ATA
Ile

Lys
AAG
AAG
Lys

Ala
GCA
GCA
Ala

Asn
AAC
AAC
Asn

Val
GTC
TTC
Phe

Leu
CTG
CTG
Leu

Leu
TTA
TTA
Leu

Thr
ACA
ACA
Thr

(c) Ile
ATA
ATA
Ile

Cys
TGT
TGT
Cys

Ile
ATA
ATA
Ile

Lys Ala Asn Val Leu Leu Thr amino acid sequence
AAG GCA AAC GTC CTG TTA ACA DNA sequence
TAG GCAAACGTCCTGTTAACA stop codon
Ter polypeptide termination

amino acid sequence

DNA sequence
base substitution
missense amino acid

Figure 2 Types of substitutions in a protein-coding region: (a) synonymous, (b) missense, and (c) nonsense. In each case, the top sequence is wild type
and the bottom sequence is mutated.

Mutation

(a) Lys Ala Leu Val Leu Leu Thr Ile Cys Ile Ter
AAG GCA CTG GTC CTG TTA ACA ATA TGT ATA TAA TACCATCGCAATAGGG
Del(G)
AAG GCA CTG vTCC TGT TAA CAATATGTATATAATACCATCGCAATAGGG
Lys Ala Leu Phe Cys Ter
(b) Lys Ala Asn Val Leu Leu Thr Ile Cys Ile Ter
AAG GCA AAC GTC CTG TTA ACA ATA TGT ATA TAA TACCATCGCAATAGGG
Ins(G)
AAG GCA AACvGvGT CCT GTT AAC AAT ATG TAT ATA ATA CCA TCG CAA AGT G
Lys Ala Asn
Gly Pro Val Asn Asn Met Tyr Ile Ile Pro Ser Gln Ter
Figure 3 Examples of frameshifts caused by deletion or insertion. (a) A deletion of a G causes premature termination. (b) An insertion of a G obliterates a
stop codon. Termination codons are underlined. In each case, the top sequence is wild type and the bottom sequence is mutated.

Insertion of transposable genetic elements

Transposable genetic elements, also designated transposons or jumping genes, are found in almost every organism.
Their most obvious behaviour is their mobility around
chromosomes. If a transposon inserts into a gene encoding
a protein, the gene is split into two pieces and the whole
protein is no longer expressed. This is the negative aspect of
transposon insertion. In contrast, there is a positive side
when, in a gene regulatory region, an inserted transposon
provides strong promoters that will increase the expression
of downstream genes.

Mutation reversions: true or pseudoreversion

of a mutation
Sometimes, after a few generations, a mutant organism
reverts to the wild type. Has the mutation reverted to the
original parental type? Not always, for the mutation may
still remain in the DNA while a secondary mutation has
occurred somewhere else, restoring a wild-type phenotype.

Mutation hotspots and mutations at

regulatory sequences
Mutation hotspots
Mutations occur, on average, randomly along the genome,
but more often than not, they crop up at some sites called
hotspots. There is such a hotspot in the Escherichia coli
chromosome: the dinucleotide CpG, in which the cytosine
is frequently methylated and replicated, with introduction
of an error, changes to TpG. Another mechanism is the
deamination of methylcytosine, which produces thymine
in the absence of replication. The dinucleotide TpT is a
4

mutation hotspot in prokaryotes (but not in eukaryotes).

Hotspots can be accounted for by the three-dimensional
shape of DNA that generates some targets for specific enzymes like methylases.
In bacteria, regions within DNA containing short palindromes, i.e. sequences that read the same on the complementary strand, such as 5GCCGGC3, 5GGCGCC3
and 5GGGCCC3, are prone to mutate more frequently
than other regions.
In eukaryotic genomes, short tandem repeats are often
hotspots, resulting from slipped-strand mispairing (Figure 4).
Mutations at regulatory sequences
DNA encodes sequences that control the expression of
proteins, such as promoters and operators. These sequences, by undergoing a mutation, may alter the cell concentration of proteins encoded downstream of the regulatory
sequence. Overexpression or underexpression of proteins
may sharply modify the function of a gene in an organism.
Since many proteins work in a concerted fashion, a sharp
increase or decrease in the cell concentration of a protein
may upset the organism.

Synonymous Mutations are Neutral

In a DNA sequence that codes for a protein, each of the
sense codons can mutate to nine other codons by means of
a single base substitution. For example, CCU, which codes
for proline (Pro), can give rise to three synonymous substitutions CCC, CCA or CCG, which also code for Pro. In
a different order, the triplet can give rise to six nonsynonymous substitutions, which code for different amino acids,

Normal pairing
of intact chromosomal DNA

Normal pairing
during DNA replication
5'__ AATCCTAGTATATA
3'
::::::::::::::
3'__TTAGGATCATATATGTGCTTAA__5'

Slipped-strand mispairing

5'__AATCCTAGTATATACACGAATT__3'
: : : : : :: : : : ::: ::: ::: : ::
3'__TTAGGATCATATATGTGCTTAA__5'

Slipped-strand mispairing

AG
T T
C A
3' 5'__AATCCTAGTATATA
5'__AATC
TATA
3'
::::
::::
::::
::::**
3'__TTAGGATCATATATGTGCTTAA__5' 3'__TTAG
ATATGTGCTTAA__5'
G
A

Replication continues
inserting TA repeat unit
AG
T T
C A

>>>>>>>>>>

Slipped-strand mispairing
AG
T T
C A
5'__AATC
TATACACGAATT__3'
::::
::::
::::
3'__TTAGGATCATAT
TTAA__5'

T
A
TC

A
T

Replication continues after

excision of unrepaired TA
repeat unit

Excision or mismatch repair inserts

TA repeat unit
AG
T T
C A

>>>>>>>>

5'__AATC
TATATACACGAATT__3' 5'__ AATCCTAGTATACACGAATT__3'
::::::::::::::
::::
: : : :: : ::: : ::
: :::
ATATGTGCTTAA__5'
3'__TTAGGATCATATATGTGCTTAA__5' 3'__TTAG
G T
A A
TC
(a)

C
G
GT

5'__AATC
TATATACACGAATT__3'
: :::
: : : : : : : : : : : :: :
3'__TTAGGATCATATATGTGCTTAA__5'

(b)
Mutation

Figure 4 Generation of duplications or deletions by slipped-strand mispairing between contiguous repeats (bold red). Small arrows indicate the direction of DNA synthesis. Dots indicate base
pairing. (a) A two-base slippage in a TA repeat during DNA replication. Slippage in the 3!5 direction results in the insertion of one TA unit (left panel). Slippage in the other direction results in the
deletion of one repeat unit (right panel). The deletion shown in the right panel results from excision of the unpaired repeat unit (asterisks) at the 3 end of the growing strands, presumably by the 3!5
exonuclease activity of DNA polymerase. (b) A two-base slippage in a TA repeat in nonreplicating DNA. Mismatched regions form single-stranded loops, which may be targets of excision or mismatch
repair. The outcome (a deletion or an insertion) will depend on which strand is excised or repaired and which strand is used as template in the DNA repair process.

Mutation

UCU for serine (Ser), ACU for threonine (Thr), GCU for
alanine (Ala), CUU for leucine (Leu), CAU for histidine
(His), or CGU for arginine (Arg).
Since the genetic code denes 61 sense codons, the possible codon substitutions can be calculated to be
61
9 5 549. If we assume that base substitutions may occur at random and that all codons are equally frequent in
regions coding for proteins, we can compute the expected
proportion of the different types of base substitutions. Because of the structure of the genetic code, synonymous
substitutions occur mainly at the third position of codons.
Indeed, 69% of all the possible synonymous changes in a
codon are at the third base position.
In contrast, substitutions at the second position of codons are nonsynonymous, and so are the vast majority of
base changes at the rst position (96%).
Synonymous amino acid replacements do not alter either protein structure or function. From an evolutionary
viewpoint, these mutations are said to be neutral, as they
do not seem to affect the tness of the mutated organism.
Such synonymous mutations accumulate with time as they
are not counterselected. Indeed, synonymous mutations
generate change at the DNA level, which has been visualized by the use of restriction enzymes and sequencing
techniques. In the long run, the accumulation with time of
synonymous mutations increases DNA divergence between individuals within a species. A new species can then
emerge by segregation. Chimpanzees and humans have
some proteins displaying a 99% similarity, but the DNA
sequences in the two species have strongly diverged, preventing the formation of hybrid species by recombination.
A species is dened by the property of the individuals
within a group to mate and give rise to progeny. At the
molecular level, it means that the two parental DNAs are
identical, so they can recombine. If genomic DNA accumulates third-base changes with time, the DNA of the
mating pair becomes too divergent with respect to their
common ancestor. The resulting diverged individuals may
attempt to mate but they cannot give rise to a fertile progeny.
Mating is always followed by DNA recombination,
which is successful only if it arises between two homologous DNAs. Mating is doomed to fail if it is forced to occur
between two heterologous DNAs. The rst step in recombination is the pairing of two DNA strands. When two
heterologous DNA strands attempt to pair, base-to-base
mismatches arise between the bases that should complement. Mismatches are removed by the induction of a very
efcient repair process called mismatch repair. It has been
shown that recombination between two heterologous
DNAs is aborted by mismatch repair.

Mechanisms Generating Mutations

Spontaneous mutagenesis
Naturally occurring mutations may arise in any cell with
no apparent cause. They result from four main mechanisms: (1) DNA polymerization errors; (2) spontaneous
oxidative DNA damage; (3) loss of a purine base, or less
frequently, loss of a pyrimidine base; and (4) deamination
of cytosine and adenine.
Polymerization errors
Point mutations, usually transitions, arise as a result of two
successive faulty processes: (1) an error in DNA replication; (2) an inefcient repair of the generated mismatches.
DNA polymerase incorporates an incorrect base at a
frequency of about 10 2 6 to 10 2 7 per base pair per cell per
generation. Normally, the incorrect base is removed by
mismatch repair, which reduces the overall rate of replication and postreplication errors down to 10 2 9 to 10 2 10
per base pair per cell per generation (frequencies observed
in E. coli).
Replication slippage, also called slipped-strand mispairing, is one of the major mechanisms that accounts for
frameshift mutations occurring at contiguous short repeats
(Figure 4).
Incorporation of an oxidized base
Exposed to oxygen, guanine is subject to oxidation, producing 8-oxo-G. This abnormal base slips into DNA, generating transversions. Fortunately, there is an elaborate
repair system that either removes 8-oxo-G or decreases the
consequences of its incorporation. Such a repair system is
ubiquitous, protecting DNA from lesions due to oxygen.
In short, the oxygen we breathe is not harmless. Oxygen
is a DNA-damaging agent and may play a role in the ageing
process. The number of oxidative hits to DNA has been
correlated with the amount of DNA breakdown products
released in rat and human urine. The number of oxidative
hits to DNA per cell per day is around 100 000 in the rat and
about 10 000 in the human. The 10-fold difference may
result in part from the greater metabolic rate of rodents,
which is at least ve times higher than that of mammals.
The mutagenic action of 8-oxo-G suggests that there is
no clear-cut border between natural and articial DNAdamaging agents. The difference between spontaneous and
induced mutations is also blurred when the air we breathe
induces the insertion of oxides into DNA.
Loss of a nucleic base
This is the most frequent spontaneously arising lesion. The
bond between sugar and base can be severed, releasing a
free base and leaving a gap in the sequence of bases, thus
generating an abasic site (apurinic or apyrimidinic). Loss

Mutation

of a purine base is more frequent than loss of a pyrimidine

base.
Replication over an abasic site may produce a point
mutation, as the polymerase has no way of determining the
identity of the missing base it has to copy. By a fortunate
trick of Nature, adenine is frequently placed opposite the
missing base. When the missing base is thymine, the placement of adenine restores the DNA code.

Deamination
This process may convert cytosine to uracil and adenine to
hypoxanthine. Conversion of cytosine to uracil is favoured
as the main cause of immunoglobin hypermutation, thus
explaining how half a million of the antibodies constituting
our immunological repertoire are made.

Induced mutations
They may result from three different mechanisms: (1) a
base in the DNA is replaced by a base analogue that confuses the polymerase; (2) a base is chemically altered so that
it mispairs with another base during replication; (3) a base
is replaced by a DNA single-strand gap so that it does not
convey any coding information. In the last case, a lowdelity replicative mechanism (SOS mutagenic repair) is
induced and bypasses the lesion.

Base replacement by a base analogue

2-Aminopurine is a base analogue because it mimics a
normal base. When incorporated into DNA, it causes frequent mispairing.

Mispairing by alkylation of bases

Alkylating agents such as ethyl-methanesulfonate and
MNNG (N-methyl-N-nitro-N-nitrosoguanidine) produce
mispairing. Although these agents modify different bases
at various positions, alkylations at the O6 position of guanine and at the O4 position of thymine cause specific mispairing with T and with G, leading to GC $ AT and AT $
GC transitions.
In vivo, mutations are mostly produced by O6-alkylguanine or O4-alkylthymine.

Noncoding lesions
Specific DNA lesions arise in DNA irradiated with ultraviolet light: pyrimidine dimers and 64 pyrimidine
pyrimidone compounds. Such lesions sharply bend
DNA, thus preventing the precise pairing of bases during
replication.

A Majority of Mutagens are

Carcinogens. The Ames Test
Geneticists consider that spontaneous mutations are relatively rare. To increase the appearance of mutations, they
have used various mutagens. We know that most mutagens
damage DNA. Bruce Ames asked an interesting question:
Do mutagens cause cancer? If they do, would it be possible
to replace the usual lengthy cancer tests on mice (lasting
months and years) with a short bacterial test?
Ames and his collaborators devised a mutagenicity test
using Salmonella tester strains (his 2 ) that require histidine
for their growth. The tester bacteria treated with a potential carcinogen were checked for the production of his+
mutations, enabling the bacteria to grow without histidine.
To be efcient, the bacterial tester strains were engineered
so as: (1) not to remove the DNA lesions produced by the
chemical tested; (2) to increase the rate of mutation by a
mutator plasmid; and (3) to render the tester strains permeable to chemical compounds. The chemical is mixed
with rat liver enzymes to simulate metabolization of the
compound by a human liver. In short, the test uses a transient physiological chimaera made of rat enzymes plus
permeable bacteria.
It was observed that, even though bacteria are evolutionarily distant from rodents and humans, they provide
an excellent tool for measuring mutagenicity and therefore
potential carcinogenicity. The results of the tests have
shown that 65% of the chemical compounds tested, which
are carcinogens in rodents, are mutagens in bacteria. The
converse is also true: most mutagens are active carcinogens. Use of the Salmonella test to detect potential carcinogens has saved time, money and suffering for animals.
Because the Ames mutagenicity test is rapid, accurate and
relatively simple, many agencies and laboratories throughout the world have tested newly produced chemicals for
their mutagenicity and potential carcinogenicity. In the
European Union, the Ames test has thus become mandatory.

Carcinogens Leave No Definite

Mutational Signatures
In human cells, a gene called p53 expresses a protein that
prevents the occurrence of cancers. Protein p53 is involved
in the control of cell division and is thus an antioncogene.
Strikingly, one nds a p53 mutation in more than half of all
cancer types taken together, 90% of which are point mutations and two-thirds of them are transitions GC ! AT.
In contrast, in bronchial cancers one nds that 45% are GC
! TA transversions. Benzopyrene, the main mutagen
present in tobacco smoke, produces GC ! TA transversions at low frequency. More generally, does the nding in
7

Mutation

cancer cells of a GC ! TA transversion in p53 prove that

benzopyrene has caused the mutation? Since a large variety
of carcinogens, including many pollutants, cause GC !
TA transversions, the answer is negative. A point mutation
is a prevalent and so minute DNA change that it cannot be
taken alone as a mark left by a carcinogen.
In order to provide legal proof, a mutated DNA sequence must be long and varied enough to be exclusive.
The probability of nding another identical copy of a given
DNA ngerprint is below 10 2 8. DNA microsatellites,
whose lengths are between 1000 and 20 000 bases, provide
valid legal proof of identication.
Only a few chemicals are recognized legally as carcinogenic. For instance, asbestos, an oxidizing mineral, causes
a unique type of tumour. Radioactive iodine ingested by
children after the Chernobyl nuclear accident has been
found to break a specific chromosome in thyroid cancer
cells. Apart from a few exceptions, there is still a debatable
causal relationship between mutation types induced by
carcinogens and specific tumours. This reality undermines
the concept that carcinogens might leave a mutational
signature on DNA.

Further Reading
Ames BN (1979) Identifying environmental chemicals causing mutations
and cancer. Science 204: 587593.
Ames BN, Shigenaga MK and Hagen TM (1993) Oxidants, antioxidants, and the degenerative diseases of aging. Proceedings of the National Academy of Sciences of the USA 90: 79157922.
Devoret R (1979) Bacterial tests for potential carcinogens. Scientific
American 241: 4050.
Echols H and Goodman MF (1991) Fidelity mechanisms in replication.
Annual Review of Biochemisty 60: 477511.
Friedberg EC, Walker GC and Siede W (1995) DNA Repair and Mutagenesis. New York: AMS Press.
Griths JF, Gelbart WM, Lewontin RC and Miller JH (2002) Modern
Genetic Analysis (Integrating Genes and Genomes). New York: Freeman.
Klein J and Takahata N (2002) Where Do We Come From? The Molecular Evidence for Human Descent. Berlin: Springer.
Li WH and Graur D (1991) Fundamentals of Molecular Evolution. Sunderland, MA: Sinauer.
Radman M and Wagner R (1988) The high delity of DNA duplication.
Scientific American 259: 4046.