Molecular mechanism of mutation by chemical mutagen

 Induced mutations are those that increase the rate of mutation above the
spontaneous background.

 fungal toxins, cosmic rays, and ultraviolet light, are natural components of our
environment.

 industrial pollutants, medical X rays, and chemicals within tobacco smoke, can be
considered as unnatural or human-made additions to our modern world.
Base Analogs

 compounds that can substitute for purines or

pyrimidines during nucleic acid biosynthesis.
 5-bromouracil (5-BU), a derivative of uracil,
behaves as a thymine analog but is
halogenated at the number 5 position of the
pyrimidine ring.
 Tautomeric shift – keto and enol form.
 If 5-BU is chemically linked to deoxyribose,
the nucleoside analog bromodeoxyuridine
(BrdU) is formed.

 If 5-BU is incorporated into DNA in place of thymine and a tautomeric shift to the enol
form occurs, 5-BU base-pairs with guanine.

 After one round of replication, an A “ T to G ‚ C transition results.

 Furthermore, the presence of 5-BU within DNA increases the sensitivity of the molecule
to ultraviolet (UV) light, which itself is mutagenic.
 2-amino purine (2-AP) can act as an analog of adenine. In addition to its base-
pairing affinity with thymine, 2-AP can also base-pair with cytosine, leading to
possible transitions from A “ T to G ‚ C following replication.
 Alkylating, Intercalating, and Adduct-Forming Agents
 A number of naturally occurring and human-made chemicals alter the structure of DNA
and cause mutations.
 sulfur-containing mustard gases, discovered during World War I.
 Mustard gases are alkylating agents—that is, they donate an alkyl group, such as CH3 or
CH3CH2, to amino or keto groups in nucleotides.
 Ethylmethane sulfonate (EMS), for example, alkylates the keto groups in the number 6
position of guanine and in the number 4 position of thymine. As with base analogs,
base-pairing affinities are altered, and transition mutations result.
 For example, 6-ethylguanine acts as an analog of adenine and pairs with thymine.
 Intercalating agents are chemicals that have dimensions and shapes that allow them to
wedge between the base pairs of DNA.
 When bound between base pairs, intercalating agents cause base pairs to distort and DNA
strands to unwind.
 changes in DNA structure affect many functions including transcription, replication, and
repair.
 Deletions and insertions occur during DNA replication and repair, leading to frame-shift
mutations.
 Example is ethidium bromide.
 Other intercalating agents are used for cancer chemotherapy. Examples are doxorubicin,
which is used to treat Hodgkin lymphoma, and dactinomycin, which is used to treat a
variety of sarcomas.
 Another group of chemicals that cause mutations are known as adduct-forming agents.

 A DNA adduct is a substance that covalently binds to DNA, altering its conformation and
interfering with replication and repair.

 acetaldehyde (a component of cigarette smoke) and heterocyclic amines (HCAs).

 HCAs are cancer-causing chemicals that are created during the cooking of meats such as
beef, chicken, and fish.

 HCAs are formed at high temperatures from amino acids and creatine.

 Many HCAs covalently bind to guanine bases. At least 17 different HCAs have been linked
to the development of cancers, such as those of the stomach, colon, and breast.
 Ultraviolet Light

 All electromagnetic radiation consists of energetic waves that we define by their different
wavelengths.

 The full range of wavelengths is referred to as the electromagnetic spectrum.

 energy of any radiation in the spectrum varies inversely with its wavelength.
 Waves in the range of visible light and longer are benign when they interact with most
organic molecules.
 energy of any radiation in the spectrum varies inversely with its wavelength
 However, waves of shorter length than visible light, being inherently more energetic, have
the potential to disrupt organic molecules.
 purines and pyrimidines absorb ultraviolet (UV) radiation most intensely at a wavelength of
about 260 nm.
purines and pyrimidines absorb ultraviolet (UV) radiation most intensely at a
wavelength of about 260 nm.
 Ionizing Radiation
 the energy of radiation varies inversely with wavelength.
 X rays, gamma rays, and cosmic rays are more energetic than UV radiation.
 As a result, they penetrate deeply into tissues, causing ionization of the molecules
encountered along the way. Hence, this type of radiation is called ionizing radiation.
 As ionizing radiation penetrates cells, stable molecules and atoms are transformed into
free radicals —chemical species containing one or more unpaired electrons.
 Ionizing radiation can directly or indirectly affect the genetic material, altering purines and
pyrimidines in DNA, breaking phosphodiester bonds, disrupting the integrity of
chromosomes, and producing a variety of chromosomal aberrations, such as deletions,
translocations, and chromosomal fragmentation.
 Although it is often assumed that radiation from artificial sources such as nuclear power
plant waste and medical X rays are the most significant sources of radiation exposure for
humans, scientific data indicate otherwise.
 Scientists estimate that less than 20 percent of human radiation exposure arises from
human-made sources.
 More than half of human-made radiation exposure comes from medical X rays and
radioactive pharmaceuticals.
 The greatest radiation exposure comes from radon gas, cosmic rays, and natural soil
radioactivity.
BEADLE AND TATUM: ONE GENE–ONE ENZYME
 Experimental Model

 chose the salmon colored bread mold Neurospora crassa.

 it can grow on medium containing only (1) inorganic salts, (2) a simple sugar, and (3)
one vitamin, biotin.
 Neurospora growth medium containing only these components is called “minimal
medium.”
 Beadle and his new collaborator, Edward Tatum, reasoned that Neurospora must be
capable of synthesizing all the other essential metabolites, such as the purines,
pyrimidines, amino acids, and other vitamins, de novo.
 Furthermore, they reasoned that the biosynthesis of these growth factors must be
under genetic control.
 mutations in genes whose products are involved in the biosynthesis of essential
metabolites would be expected to produce mutant strains with additional growth
factor requirements.
Beadle and Tatum tested this prediction by irradiating asexual spores (conidia) of wild-
type Neurospora with X rays or ultraviolet light, and screening the clones produced by
the mutagenized spores for new growth-factor requirements
 By correlating their genetic analyses with biochemical studies of the mutant strains,
they demonstrated in several cases that one mutation resulted in the loss of one
enzyme activity.

 Beadle and Tatum received a Nobel Prize in 1958.

 One-Gene-One-Enzyme / One-Gene-One-Polypeptide Concept

 E. coli, the enzyme tryptophan synthetase is a heterotetramer composed of two

polypeptides encoded by the trpA gene and two polypeptides encoded by the trpB
gene.

 Hemoglobins, which transport oxygen from our lungs to all other tissues of our bodies.

 tetrameric proteins that contain two -globin chains and two -globin chains, as well as
four oxygen-binding heme groups.

 E. coli DNA polymerase IIIand RNA polymerase II, contain many different polypeptide
subunits, each encoded by a separate gene.

 Thus, the one gene–one enzyme concept was modifi ed to one gene–one polypeptide.