Mutagen

A mutagen is a physical or chemical agent that has the ability to cause mutations in DNA and raises
their frequency above natural background levels.
They may directly damage DNA as a result of their actions, which most frequently leads to
replication errors.

Mutations occur in a random manner, i.e., they are not directed according to the requirements of the
organism. The majority of mutations are caused by environmental factors, but they can also be
created in the lab using chemicals, radiation, or physical stimuli.

These mutagens may be carcinogens because numerous mutations can lead to cancer in animals,
though not necessarily in all cases.
All mutagens exhibit distinctive mutational signatures, while some chemicals can become mutagenic
as a result of cellular processes.

Types
Mutagens can be classified into 3 major types based on its origin. They are:
1. Physical mutagen
2. Chemical mutagen
3. Biological mutagen

Physical Mutagen
It includes:
i. High energy radiations
ii. Increase in temperature

i. Radiation
The radiations which are important in mutagenesis are of two categories. They are:
A. Ionizing radiations: X-rays and gamma rays; alpha and beta rays; electrons, neutrons,
protons and other fast moving particles
B. Non-ionizing radiations: Ultraviolet and visible light.
A. Ionizing radiations
The process by which ionizing radiations trigger mutation is still mostly unknown. Ionizing
radiation causes chromosomal changes such break, deletion, addition, inversion, and
translocation by damaging the poly sugar phosphate backbone of DNA.
The active function of oxygen is anticipated during DNA molecule breaking caused by ionizing
radiation. Because oxygen is necessary for the production of H2O2 and HO2, which may cause
DNA molecule breakage in irradiated water.
The most common lab sources include cobalt-60 and cesium-137.

B. Non-ionizing radiations
The ultraviolet (UV) light is a non-ionizing radiation which may cause mutation.
The most effective wave length of ultraviolet for inducing mutations is about 2,600 A°. This is a
wave length that is best absorbed by DNA and a wave length at which proteins absorb little
energy.
Some of the ways that UV radiations induce mutations include base deletion, strand breakage,
cross-linking, and the formation of nucleotide dimers.
UV radiations are of three types:
UV-A has a wavelength of 320nm (near-visible range) and is known to cause dimerization of
pyrimidines. This particular form of pyrimidine dimerization alters the DNA structure,
preventing the creation of the replication fork during the replication process. Such dimerization
could have negative health effects.
UV-B has a wavelength of 290-320nm and highly lethal to DNA.
UV-C has a wavelength of 180-290nm which is majorly absorbed by the ozone layer and are the
most lethal as well as carcinogenic.

ii. Temperature
The rate of all chemical reactions are influenced by temperature.
A temperature increase of 10°C doubles or triples the rate of mutation.
The thermal stability of DNA and the rate at which other substances react with it are both
impacted by temperature, which also destroys the hydrogen and phosphodiester bonds that are
present in DNA.
Chemical Mutagen
Many chemical compounds are known to increase the mutability of genes.
Auerbach and Robson’s experiment with male Drosophila melanogaster in 1947, utilizing mustard
gas and related substances such as the nitrogen and Sulphur mustards, mustard oil, and chloracetone,
were the first to show that chemicals can cause mutations.
Chemical mutagens affect the chromosomal DNA by following two ways:
1. Direct gene change
2. Copy error

Chemical mutagens can be classified into different categories such as:

i. Base analogs
ii. Intercalating agents
iii. Metal ions
iv. Alkylating agents

i. Base analogs
These substances have structural characteristics with bases like purines and pyrimidines.
5-Bromouracil and aminopurine are the two most prevalent base analogs that are considered to be
chemical mutagens.
Base analogs are integrated into the DNA structure during replication because of the structural
resemblances between these agents and DNA bases.
Similar to adenine, aminopurine can pair up with either C or T to form a base pair (though base
pairing with C is rare).
Some additional base analogues, including urethane triazine, caffiene (found in coffee, tea, and
soft beverages), phenol and carcinogens, acridines (proflavin, etc.), and others, are similarly
mutagenic.

ii. Intercalating agents

Intercalating molecules are those with a hydrophobic heterocyclic ring structure that resembles
the base pair ring structure.
These agents embed themselves in the DNA helix, causing interference with transcription,
replication, and mutation, most frequently a frameshift mutation.
 Some of the common intercalating agents are:
 Ethidium bromide
 Proflavine
 Acridine orange
 Actinomycin D
 Daunorubicin

iii. Metal ion

Reactive oxygen species (ROS) are produced by mineral ions such as nickel, chromium, cobalt,
cadmium, arsenic, chromium, and iron that lead to DNA hypermethylation. This promotes DNA
damage and obstructs the DNA repair process.
For many organisms, certain inorganic chemicals like manganese chloride are mutagenic because
they bind calcium and interfere with the integrity of the chromosome structure.

iv. Alkylating agents

These chemicals cause DNA damage by inducing alkyl groups.
Alkyl group introduction boosts ionization, leading to base-pairing mistakes that eventually cause
holes in the DNA strand and have a direct mutagenic effect on the DNA molecule.
Some of the common alkylating agents are:
 Nitrous acid
 Ethylnitrosourea
 Methylhydrazine
 Dacarbazine
 Formaldehyde
 Vinyl chloride
 Epoxides
 Dimethyl and diethyl sulphonate
 Methyl and ethyl methanesulphonate (MMS and EMS)
 Nitrosoguanadine (NG)
However, these substances can be eliminated from the DNA by the depurination process during
the DNA repair process.
Biological Mutagen
It includes:
i. Transposons and Insertion sequences (IS)
ii. Viruses
iii. Bacteria

i. Transposons and Insertion sequences (IS)

They are DNA units that carry out the DNA fragment’s self-directed relocation and
multiplication.
Insertion sequences, or IS, are the shortest type of transposons (between 10 and 50 base pairs
long).
Since IS and transposon migrate around the DNA, they are both referred to as jumping genes.
The functionality of the genes is disrupted when transposons are inserted into chromosomal
DNA.
Information for the enzyme transposases, which aids in creating the new transposition location in
the DNA, is encoded by both IS and transposons.
Three types of transposons are usually found. They are:
A. Replicative transposons: The transposons that retain the original locus and translocate its
copy.
B. Conservative transposons: The original transposon translocates itself.
C. Retrotransposons transpose: These transposons translocate via RNA intermediates.

ii. Viruses
The insertion of viral DNA into the genome may lead to the disruption of genetic function.
The viruses cause deletions, insertions, and point alterations, including base substitutions.
The stimulation of cells’ prone-to-error repair mechanisms is reported to be connected to virus-
induced mutagenesis.
Rous sarcoma virus have been reported to induce cancer.

iii. Bacteria
Reactive oxygen species are produced by some bacteria that cause inflammation, such as
Helicobacter pylori, leading to DNA damage and decreased DNA repair.
By boosting activation-induced cytidine deaminase (AID), a DNA/RNA editing enzyme that
links mutagenesis and cancer, H. pylori impacts genome integrity.
Positive effects of Mutagen
1. Evolution
The changes in the gene pool that ultimately contributed to the evolution of life over time are
caused by mutations.
Natural selection has allowed populations with a variety of mutations to persist, while
populations that were unable to adapt to environmental changes or evolve as a result of those
changes eventually perish.
One such genetic evolutionary stage involves the development of coat color by insects and
animals for camouflage.

2. Adaptation
Apolipoprotein A1-Milano (or Apo A1M), a mutant protein, is found in a small Italian group.
Normal Apolipoprotein is the protein responsible for the transportation of cholesterol.
The mutant form of apolipoprotein, possesses antioxidant characteristics in addition to removing
cholesterol and dissolving plaques and has aided the Italian population from cardiac diseases.

3. Antibiotic Resistance in Bacteria

Mutations in many bacteria enable them to endure the presence of antibiotic medications. The
mutations result in bacterial strains that are resistant to antibiotics.

Negative effects of Mutagen

1. Sickle cell anemia, which is brought on by a single missense mutation in the germ cell -globin
gene at codon 6. With this mutation, valine takes the place of glutamic acid at position 6 in the
original protein. Hemoglobin, a protein that transports oxygen, is negatively impacted by this
alteration.
2. Some pesticides, including rotenone, paraquat, and maneb, have the potential to cause mutations
in genes, such as base pair alterations, which may result in neurological illnesses like Parkinson
disease.
3. Retinoblastoma or Retinal tumors in children.
4. Genetic disorders such as Tay-Sachs disease, Phenylketonuria, Color-blindness, and Cystic
fibrosis
Mutagenicity Testing
Identifying compounds that potentially change the genetic makeup of somatic and/or germ cells is
the objective of mutagenicity testing, which is used to influence regulatory decisions.
Genetic alterations are usually manifested after a long time after exposure to mutagen, in contrast to
the majority of other types of toxicity.
Three endpoints are usually assessed to provide an adequate evaluation of the mutagenic potential.
They are:
1. Gene mutation
2. Structural chromosome aberrations
3. Numerical chromosome aberrations

The commonly used in vitro and in vivo studies are:

In Vitro Studies
1. Bacterial reverse mutation test - Ames test
2. Mammalian chromosome aberration test
3. Mammalian cell gene mutation test
4. UDS in mammalian cells

In Vivo Studies
1. Rodent dominant lethal mutation test
2. Mouse heritable translocation assay
3. Mouse-specific locus test
4. Sister chromatid exchange (SCE) analysis in spermatogonia
5. UDS test in testicular cells
6. Transgenic rodent somatic and germ cell gene mutation assay
7. Mammalian erythrocyte micronucleus test
Antimutagen
It is an agent capable that has the ability to inactivate a mutagen, stop the mutagen from acting, or
otherwise prevent a mutagen’s interaction with DNA.
It may directly or indirectly trigger, inhibit, or inactivate the enzymes of the DNA repair,
recombination, and replication pathways.
The antimutagens can be classified as:
1. Desmutagens
2. Bio-antimutagens

1. Desmutagens
These are compounds that, through enzymatic or chemical interaction, partially or completely
render mutagens inactive before the mutagen affects the genes.
2. Bio-antimutagens
These are regarded as true antimutagen which suppress the mutation after genes are damaged by
mutagens. The mutagen-damaged DNA is repaired and replicated by them, which reduces the
frequency of mutations.

Mechanism of Antimutagenesis
The major mechanisms of antimutagenesis can be broadly described as under:
1. Chemical or enzymatic inactivation
2. Prevention of formation of active species
3. Scavenging
4. Antioxidant free radical scavenging

1. Chemical or enzymatic inactivation

Numerous mutagens can be directly inactivated by a variety of different substances since they are
reactive and act not only on DNA but also on proteins and enzymes.
Both inducers and inhibitors of cytochrome P-450 enzymes, including indole-3-carbinol, have
been linked to antimutagenic and anticarcinogenic activities.
Example:
Isothiocyanates such as benzyl isothiocyanate
Antioxidants such as 2, 3-tert butyl-4-hydroxy-anisole (BHA).
2. Prevention of formation of active species
Numerous genotoxic mutagens or carcinogens need to be metabolically or biologically activated
before they may become an electrophilic species (the active species) and interact with DNA.
Although these processes frequently take place in the liver, there is mounting evidence that other
organs, particularly the GIT, can also activate metabolism.
Example:
N-nitro compounds frequently result from an interaction between secondary or tertiary amines
and nitrite in the stomach.

3. Scavenging
Several desmutagens can bind to or adsorb dietary mutagens in order to remove them.
The mutagen often does not change during this phase but is unable to interact with DNA.
Example:
Chlorophyllin and some dietary fibers.

4. Antioxidant and free radical scavenging

Free radicals can harm DNA, lead to mutagenicity and cytotoxicity, and as a result, are crucial to
the development of cancer.
reactive oxygen species (ROS) can cause mutations and prevent DNA repair, which inactivates
some tumor suppressor genes and results in cancer.
Numerous antimutagenic substances have antioxidant or free radical scavenging properties and
are able to quickly neutralize the majority of free radicals, especially those with a short half-life.
Example:
OH radical.
Antimutagenic compounds
The major classes of antimutagenic compounds include:

i. Vitamins:
 Vitamins C and E are found to be antimutagenic against doxorubicin induced chromosomal
aberrations.
 Vitamins A, C, and E were reported to be antimutagenic towards Methyl Azoxy Methanol
(MAM) induced mutagenesis in Salmonella Typhimurium strain TA100.
 When given with a pesticide, vitamin C (ascorbic acid) significantly reduced the frequency of
pesticide-induced mutations.

ii. Flavonoids
 Leuteolin, kaempherol, and other phenolic hydroxyl group-containing flavonoids, as well as all
flavones, exhibited antimutagenic properties.
 Isoflavones and flavonoid glycosides have been found to have potent antimutagenic properties.
 Hispidulin and hortensin were found to have an antimutagenic impact when tested against 2-
amino anthracene, a compound that causes mutations caused by aflatoxin B1.
 Citrus juice flavonoids have anticarcinogenic and antimutagenic qualities.
 Salmonella Typhimurium has been used to investigate the antimutagenic activity of all these
flavonoids against various types of mutagens.

iii. Anthraquinones
 Aloe barborescence anthraquinones have been found to have antimutagenic properties.
 Anthrone, acridone, and xanthone, chemicals chemically similar to anthraquinones, exerted
antimutagenecity, with anthrone being the most potent example.
 All naphthaquinones are potent antimutagens.
iv. Phenolic compounds
 Certain phenolic chemicals, such as ellagic acid, which is present in strawberries, raspberries,
grapes, walnuts, etc., are reported to be antimutagenic.
 The antimutagenic properties of green tea and black tea have been attributed to substances like
epicatechin, (-) epicatechin gallate, (-) epigallocatechins, and (-) epigallocatechin gallate.
 Green tea's antimutagenic properties can stop the frequency of sister chromatid exchanges
induced by smoking cigarette,
 The phenolics present in turmeric and clove, namely curcumin and eugenol, were found to inhibit
the mutagenicity produced by direct acting mutagens such as N-methyl-N'-nitro-N-
nitrosoguanidine. 
 Eugenol inhibits tobacco-induced mutagenesis in the Ames test. 

v. Carotenoids
 Carotenoids are known to affect activation of promutagens.
 Antimutagenic activity of β-carotenre, canthaxanthin, β-carotene-8-apo-β-carotenal and 8-apo-β-
carotene methyl ester showed a dose dependent decrease in the mutagenicity.

vi. Diterpenoids
 Erythroxydiol, a diterpenoid-like compound isolated from Aquillaria agallocha, exhibited
antimutagenic and anticancer action.
 Pulcherrimins A, B, C, and D, four new dibenzoate diterpenes isolated in the roots of
Caesalpinia pulcherrima, were discovered to be active in a DNA repair-deficient yeast strain.

vii. Coumarins
 Antimutagenic compounds such as coumarin, umbelliferone, and 8-methoxypsoralen have been
identified from various plant sources.
 Psoralen from Psoralea corylifolia, imperatorin, and osthol from Selinum monniere can reduce
the mutagenicity brought on by benzo[α]pyrene.
viii. Tannins
 Some tannins’ antioxidative properties are crucial in preventing cellular oxidative damage such
lipid peroxidation, to their anticarcinogenic and antimutagenic capabilities.
 Catechin, ellagic acid, and gallic acid have antimutagenic properties against mutagenicity
brought on by known mutagens.

ix. Hormonal steroid

 The synthetic β-oestradiol derivatives ethinyl oestradiol and mestranol, which are both
commonly found in birth control tablets, were both potent mutagenicity inhibitors that acted at
nanomolar doses.
 The hormones testosterone, β-oestradiol, and dicthyl stilbesterol were antimutagenic and co-
recombinogenic in experiments employing yeast without an external metabolic activation
mechanism.

x. Saponins
 Human gut bacteria-introduced ginseng saponin metabolites were discovered to be antigenotoxic
against benzo[α]pyrene-induced clastogenecity.

xi. Marine products

 A variety of mutagens have the potential to be inhibited in their ability to cause mutations in
Salmonella Typhimurium by certain secondary metabolites prevalent in marine species.
 Elatol and obtusol are two antimutagenic substances that were discovered in a sea hare extract.
 By using an antimutagenicity assay, the green algae Cymopolia barbata was used to isolate
cymobarbatol and 4-isocymobarbatol.

xii. Miscellaneous compounds
 The organosulfur compounds present in garlic extract with considerable antimutagenic activity
include ajoene and one of allicin's derivatives.
 Numerous other diverse phytocompound subgroups, including caffeine, trigonelline, and
piperine, have been shown to have antimutagenic activities.
 When isolated from Visma amazonica, xanthones such euxanthone and 1,5-dihydroxy-8-
methoxyxanthone exhibit significant antimutagenic action against 2-aminoanthracene and EMS.
 In a Salmonella mutation assay, an 80% ethanol extract of lemon grass (Cymbopogon citrates)
was discovered to be antimutagenic against a number of recognized mutagens.
 Lemongrass essential oil have antimutagenic effect against lead nitrate and cyclophosphamide-
induced micronuclei and chromosomal aberration. This oil is utilized as a component of
Lemongrass Tea in central and southern India.
Food Products as Antimutagens
 Dietary desmutagens function as tumor growth inhibitors later in the carcinogenesis process.
 Some regularly ingested spices and vegetables, including turmeric, mustard, green leafy
vegetables, and allium species, have been found to have antimutagenic and anticarcinogenic
properties.
 A human intervention trial using various carotenoids in vegetable products indicated that adding
tomato, carrot, or spinach products to the diet significantly reduced the amount of lyphocyte
DNA damage.
 Using the Salmonella Typhimuium strains TA100 and TA1535, the garlic extract inhibited the
mutagenicity caused by direct acting mutagens such N-methyl-N'-nitro-N-nitrosoguanidine and
sodium azide.
 In the DNA repair host mediated assay and liquid suspension experiment, casein demonstrated a
high antimutagenic action both in vivo and ex vivo.
 Yogurt and Guava (Psidium guajava) are reported to be antimutagenic.
 The mechanism of antimutagenic effects of mushrooms was found to be by direct chemical
interaction with the mutagens viz. aflatoxin B1, benzo[α]pyrene and acridine or inhibition of the
activation process in the case of promutagens.
 Asafoetida and turmeric extracts suppress microsomal activation dependent mutagenicity of 2-
acetamido fluorine.
 Indian spinach leaf extract; curcumin and eugenol which are phenolics present in turmeric and
clove, are reported to be antimutagenic.
 In the Ames test, alkyl-resorcinols, amphiphillic substances frequently present in wheat grains,
showed evidence of antimutagenicity.
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