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5ʹ–AACGCTAGATC–3ʹ 5ʹ–AACGCTC–3
→
3ʹ–TTGCGATCTAG–5ʹ 3ʹ–TTGCGAG–5ʹ
Gene Mutations
• Addition of 4 bp
• 5ʹ–AACGCTAGATC–3ʹ 5ʹ–AACAGTCGCTAGATC–3’
→
3ʹ–TTGCGATCTAG–5ʹ 3ʹ–TTGTCAGCGATCTAG–5
Gene Mutations Can Alter Coding Sequence
Within a Gene
• Silent mutations are those that do not alter amino acid sequence of
the polypeptide even though nucleotide sequence has changed
• Genetic code is degenerate, silent mutations can occur in certain
bases within a codon, such as the third base, so the specific amino
acid is not changed
• In contrast, missense mutations are base substitutions in which an
amino acid change does occur.
• sickle cell anemia
• a mutation in the β-globin gene, alters sixth amino acid is changed
from a glutamic acid to valine
• Single amino acid substitution alters the structure and function of the
hemoglobin protein
• As a result red blood cells assume a sickle shape under conditions of
low oxygen
• Nonsense mutations involves change from a normal codon to a stop codon
• It terminates translation of polypeptide earlier than expected
• Producing a truncated polypeptide
• A nonsense mutation occurs in a bacterial operon, it may also inhibit the
expression of downstream genes, termed polarity
• Frameshift mutations involve addition or deletion of a number of
nucleotides that is not divisible by three
• Codons are read in multiples of three, this shifts the reading frame
• Translation of mRNA results in a completely different amino acid sequence
downstream from mutation
• New mutations are more likely to produce polypeptides that have
reduced rather than enhanced function
• Nonsense mutations produce polypeptides that are substantially
shorter
• Shorter polypeptides are unlikely to function properly
• Frameshift mutations dramatically alter the amino acid sequence of
polypeptides and are thereby likely to disrupt function
• Missense mutations are less likely to alter function because they
involve a change of a single amino acid within polypeptides that
typically contain hundreds of amino acids.
• Neutral mutation If a missense mutation has no detectable
effect on protein function
• A missense mutation that substitutes an amino acid with a
similar chemistry as the original amino acid is likely to be neutral or
nearly neutral
• For example, a missense mutation that substitutes a glutamic acid for
an aspartic acid is likely to be neutral
• As both amino acids are negatively charged and have similar side
chain structures
• Silent mutations are also considered a type of neutral mutation
Point mutations can alter mRNA splicing
• Single-base-pair changes which inactivate proteins are due to splice
site mutations
• It will dramatically change RNA transcript by leading to large
insertions or deletions
• Neutral mutation does not alter protein function
• Does not affect survival or reproductive success.
• Deleterious mutation, decreases chances of survival and
reproduction
• Extreme example of a deleterious mutation is a lethal mutation
• Results in death to a cell or organism
• Beneficial mutation enhances the survival or reproductive success
• An allele may be either deleterious or beneficial depending on
genotype and/or environmental conditions (sickle cell allele)
• In homozygous state, sickle cell allele lessens chances of survival
• In heterozygous (sickle cell allele and wild-type allele ) has an
increased chance of survival due to malarial resistance
Molecular Basis of Spontaneous Mutations
• Gene mutations can arise spontaneously or they can be induced
• Spontaneous mutations are naturally occurring mutations
• Induced mutations arise through the action of mutagens
• They increase rate at which mutations occur
• Salvadore Luria and Max Delbrück were interested in the
ability of bacteria to become resistant to infection by a bacteriophage
called T1.
• When a population of E. coli cells is exposed to T1, a small percentage of
bacteria are found to be resistant to T1 infection and pass this trait to their
progeny.
• Luria and Delbrück were interested in whether such resistance, called tonr
(T one resistance), is due to the occurrence of random mutations
or whether it is a physiological adaptation that occurs at a low rate within
the bacterial population
• Do spontaneous mutations occur in response to a selecting agent or
are variants present at a low frequency in most populations?
• Ideal experimental system to address this important question was
analysis of mutations in bacteria
• Luria and Delbrück fluctuation test
• if E. coli bacteria are spread on a plate of nutrient medium in the
presence of phage T1, phages soon infect and kill bacteria
• Rarely, but regularly, colonies were seen that were resistant to phage
attack
• These colonies were stable and so appeared to be genuine mutants
• Whether these mutants were produced
A- Spontaneously
B-Phage induced a physiological change that caused resistance
• If mutations occurred spontaneously, then mutations may occur at
different times in different cultures (Lauria)
• So numbers of resistant colonies per culture should show high
variation (fluctuation)
Fluctuation test
• 20 small cultures were inoculated, each with a few cells, and
incubated till 108 cells per millilitre
• At same time, a much larger culture also was inoculated and
incubated until 108 cells per millilitre
• 20 individual cultures and 20 samples of same size from large culture
were plated in presence of phage
• 20 individual cultures showed high variation in number of
resistant colonies: 11 plates had 0 resistant colonies, and remainder
had 1, 1,3, 5, 5, 6, 35, 64, and 107 per plate
• 20 samples from large culture showed much less variation from plate to
plate, all in the range of 14 to 26
• If phages were inducing mutations, there was no reason why fluctuation
should be higher on the individual cultures because all were exposed to
phage similarly
• Best explanation was that mutation was occurring randomly in time
• Early mutations gave the higher numbers of resistant cells because the
mutant cells had time to produce many resistant descendants
• Later mutations produced fewer resistant cells
• It led to the reigning “paradigm” of mutation
• “ Whether in viruses, bacteria, or eukaryotes, mutations can occur in
any cell at any time and their occurrence is random”
• It was suggested that resistant cells are selected by the environmental
agent (phage) rather than produced by it
• Luria and Delbrück were awarded the Nobel Prize in Physiology or
Medicine in 1969
• Interestingly, this was after Luria’s first graduate student, James
Watson, won his Nobel Prize (with Francis Crick in 1964) for discovery
of the DNA double-helix structure
8 bp 4-5 bp