Mutations and its types

The term mutation refers to both (1) the change in the genetic material and (2) the process by which the

change occurs. Mutations alter the nucleotide sequences of genes in several ways, for example the substitution

of one base pair for another or the deletion or addition of one or a few base pairs.

Used in its broad historical sense, mutation refers to any sudden, heritable change in the genotype of a

cell or an organism. Mutational changes in the genotype of an organism include changes in chromosome

number and structure, as well as changes in the structures of individual genes. Mutations that involve changes at

specific sites in a gene are referred to as point mutations. They include the substitution of one base pair for

another or the insertion or deletion of one or a few nucleotide pairs at a specific site in a gene. Today, the term

mutation sometimes is used in a narrow sense to refer only to changes in the structures of individual genes.

Types of Mutations

There are three types of DNA Mutations: base substitutions, deletions and insertions.

1. Base Substitutions
Single base substitutions are called point mutations, recall the point mutation Glu -----> Val which

causes sickle-cell disease. Point mutations are the most common type of mutation and there are two types.

Transition:

This occurs when a purine is substituted with another purine or when a pyrimidine is substituted with

another pyrimidine.

Transversion:

When a purine is substituted for a pyrimidine or a pyrimidine replaces a purine.

codon (both codons code for same amino acid) will be generated. Thus the amino acid sequence encoded by the

gene is not changed and the mutation is said to be silent.

Missence: When base substitution results in the generation of a codon that specifies a different amino acid and

hence leads to a different polypeptide sequence. Depending on the type of amino acid substitution the missense

mutation is either conservative or nonconservative. For example if the structure and properties of the substituted

amino acid are very similar to the original amino acid the mutation is said to be conservative and will most

likely have little effect on the resultant proteins structure / function. If the substitution leads to an amino acid

with very different structure and properties, the mutation is nonconservative and will probably be deleterious

(bad) for the resultant proteins structure / function (i.e. the sickle cell point mutation).

Nonsense: When a base substitution results in a stop codon ultimately truncating (stopping translation at the

mutated region) translation and most likely leading to a nonfunctional protein.

2. Deletions
A deletion, resulting in a frameshift, results when one or more base pairs are lost from the DNA (see

Figure above). If one or two bases are deleted the translational frame is altered resulting in a garbled message

and nonfunctional product. A deletion of three or more bases leave the reading frame intact. A deletion of one

or more codons results in a protein missing one or more amino acids. This may be deleterious or not.

3. Insertions
The insertion of additional base pairs may lead to frameshifts depending on whether or not multiples of

three base pairs are inserted. Combinations of insertions and deletions leading to a variety of outcomes are also

possible.
Causes of Mutations

1. Errors in DNA Replication

On very, very rare occasions DNA polymerase will incorporate a noncomplementary base into the

daughter strand. During the next round of replication the missincorporated base would lead to a mutation. This,

however, is very rare as the exonuclease functions as a proofreading mechanism recognizing mismatched base

pairs and excising them.

2. Errors in DNA Recombination

DNA often rearranges itself by a process called recombination which proceeds via a variety of

mechanisms. Occasionally DNA is lost during replication leading to a mutation.

3. Mutagens (mutagenic agents)

Chemical mutagens

Many chemical mutagens, some exogenous, some man-made, some environmental, are capable of

damaging DNA. Many chemotherapeutic drugs and intercalating agent drugs function by damaging DNA.

i. Base Analogues:

A base analogue is a chemical compound similar to one of the four bases of DNA. It can be incorporated

into a growing polynucleotide chain when normal process of replication occurs.’ These compounds have base

pairing properties different from the bases. They replace the bases and cause stable mutation.

A very common and widely used base analogue is 5-bromouracil (5-BU) which is an analogue of

thymine. The 5-BU functions like thymine and pairs with adenine. 5-BU base pairs with Guanine also.

Thus, it initiates AT→GC pair conversion during replication. Also it converts a GC→AT.
 The 2-amino-purine (2-AP) and 2, 6-di-amino-purine (2, 6-DAP) are the purine analogues. The 2-AP

normally pairs with thymine but it is able to form a single hydrogen bond with cytosine resulting in transition of

AT to GC. The 2-AP and 2, 6-DAP are not as effective as 5-BU and 5-BDU.

ii. Chemicals Changing the Specificity of Hydrogen Bonding:

There are many chemicals that after incorporation into DNA change the specificity of hydrogen -

bonding. Those which are used as mutagens are nitrous oxide (HNO2), hydroxylamine (HA) and ethyl-methane-

sulphonate (EMS).

(a) Nitrous Oxide (HNO2):

Nitrous oxide converts the amino group of bases into keto group through oxidative deamination. The

order of frequency of deamination (removal of amino group) is adenine > cytosine > guanine.

(b) Deamination of Adenine:

Deamination of adenine results in formation of hypoxanthine, the pairing behaviour of which is like

guanine. Hence, it pairs with cytosine instead of thymine replacing AT pairing by GC pairing.

(c) Deamination of Cytosine:

Deamination of cytosine results in formation of uracil by replacing – NH2 group with -OH group. The

affinity for hydrogen bonding of uracil is like thymine; therefore, C-G pairing is replaced by U-A pairing.

(d) Deamination of Guanine:

Deamination of guanine results in formation of xanthine, the later is not mutagenic. Xanthine behaves

like guanine because there is no change in pairing behaviour. Xanthine pairs with cytosine. Therefore, G-C

pairing is replaced by X-C pairing.

(e) Hydroxylamine (NH2OH):

It hydroxylates the C4 nitrogen of cytosine and converts into a modified base via deamination which

causes to base pairs like thyamine. Therefore, GC pairs are changed into AT pairs.
 iii. Alkylating Agents:

Addition of an alkyl group to the hydrogen bonding oxygen of guanine (N7 position) and adenine (at

N3 position) residues of DNA is done by alkylating agents. As a result of alkylation, possibility of ionization is

increased with the introduction of pairing errors. Hydrolysis of linkage of base-sugar occurs resulting in gap in

one chain.

This phenomenon of loss of alkylated base from the DNA molecule (by breakage of bond joining the

nitrogen of purine and deoxyribose) is called depurination. Depurination is not always mutagenic. The gap

created by loss of a purine can effectively be repaired.

Following are some of the important widely used alkylating agents:

(a) Dimethyl sulphate (DMS)

base (A,T,G,C) may be inserted in the gap. During replication chain without gap will result in normal DNA. In

the second round of replication gap is filled by suitable base.

If the correct base is inserted, normal DNA sequence will be produced. Insertion of incorrect bases

results in transversion or transition mutation. Another example is methyl nitrosoguanidine that adds methyl

group to guanine causing it to mispair with thyamine. After subsequent replication, GC is converted into AT

transition.

iv. Intercalating Agents:

There are certain dyes such as acridine orange, proflavine and acriflavin which are three ringed

molecules of similar dimensions as those of purine pyrimidine pairs. In aqueous solution these dyes can insert

themselves in DNA (i.e. intercalate the DNA) between the bases in adjacent pairs by a process called

intercalation.

Therefore, the dyes are called intercalating agents. The acridines are planer (flat) molecules which can

be intercalated between the base pairs of DNA; distort the DNA and results deletion or insertion after
replication of DNA molecule. Due to deletion or insertion of intercalating agents, there occur frameshift

mutations.

Physical Mutagens:

i. Radiations as Mutagens:

Radiation is the most important among the physical mutagens. Radiations damaging the DNA molecules

fall in the wavelength range below 340 nm and photon energy above 1 electro-volt (eV). The destructive

radiation consists of ultraviolet (UV) rays, X-rays, ү-rays, alpha (α) rays, beta (β) rays, cosmic rays, neutrons,

etc.

Mutants

Geneticists often distinguish between the genotype and phenotype of an organism. Strictly speaking, the

entire set of genes carried by an individual is its genotype, whereas the function and physical appearance of an

individual is referred to as its phenotype.

An organism that exhibits a novel phenotype resulting from a mutation is called a mutant.

Mutant types
Morpological Mutant.

Mutation results in change in the physical properties of an organism, such as shape, color or size.

Albino ascospores in Neurospora, curly wings in Drosophila, and dwarf peas are all morphological mutations.

Lethal Mutataions.

The mutation results in death of the cell or organism are called as lethal mutations. Certain blood

abnormalities, death of organ cells, death of organism and etc.

Which make the mutant is restricted to survive or function under specific environmental conditions only

(Restrictive condition). But the wild type of that mutant survives and functions in some different environment

conditions (Permissive condition). Example temperature sensitive mutants, conversion of pathogenic state to

non pathogenic state, pH sensitive mutants, oxygen sensitivity changes and etc. Wild type organisms from

permissive conditions of environmental growth factors shift to restrictive environmental conditions after

mutation.

Biochemical mutants

Microbial cultures are convenient material for the study of biochemical mutations, which are identified

by the loss or change of some biochemical function of the cells. This change typically results in an inability to

grow and proliferate. In many cases, however, growth of a mutant cell can be restored by supplementing the

growth medium with a specific nutrient.

Loss-of-function mutants.

Because mutation events introduce random genetic changes, most of the time they result in loss of

function. The mutation events are like bullets being fired at a complex machine; most of the time they will

inactivate it. That results in loses of some functions in the cell or organism. Generally, loss-of-function (null)

mutations are found to be recessive.

Gain of function Mutants

Rarely mutation results in production of some new function or improved activity to the cell or organism.

So mutants gain some functions or activity which is not actually present in wild type.