Chapter Seven
Mutation
Objectives
At the end of this chapter, students will be able to:
List the various types of genetic variants
Describe essential concepts of genetic variation a&
chromosome structure, & DNA sequence
List causes of induced mutation
List & explain the types of DNA damages which occur
Explain various mechanisms for DNA repair
List the pathologic manifestations of defective DNA
repair
Describe diseases related to DNA Repair deficiency
Outline
Types of mutation
Mutagenic agents
How mutation affects the genetic code?
Repair of Mutation
Mutant isolation & detection
Introduction
Mutation is a permanent change in the
sequence of nucleotides in DNA
Passed to all daughter cells (inherited)
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Types of Genetic Variants
Single gene mutation
Chromosomal mutation
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Single gene mutation
Mutation that affect only a single gene due
to changes in a nucleotide sequence
Not microscopically observable
Centers on changes in coding DNA or in the
regulatory sequences, changes in the other
parts usually have no clinical consequences
Types of single gene mutation
1. Point mutation - base pair substitution
it is mutation in which one bp is replaced
by another
replacement of one nucleotide by
another may or may not affect amino acid
sequence
consists of: silent & non-silent
Types of Point mutation
a. Silent mutation
o new codon still codes for same amino acid,
because of redundancy of genetic code,
b. Non-silent mutation: involves
Mis-sense mutation:
None sense mutation:
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Types of non-silent
Mis-sense mutation:
- codon changed such that it encodes a
new amino acid
- E.g. sickle cell anemia: glutamic acid
(normal) changed to valin (abnormal)
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None sense mutation
codon changed to stop codon; causes
early chain termination in mRNA
resulting Incomplete (nonsense protein
chain) (inactive products).
Conversely. If the stop codon is altered
it encodes an abnormally elongated
polypeptide can
2 Frame shift mutation
Addition or deletion of one or more base
pairs
This adding or deleting a single bp, change
the reading frame of the transcribed mRNA
Result in frame-shift mutations, & different
polypeptides being made, usually
nonfunctional
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Chromosomal mutation
1. Chromosome number abnormality
Each organism has a typical number of
chromosomes
Each organism has a typical number of
sets of chromosomes
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Chromosome number abnormality
A. Euploidy:
the chromosome number is a multiple of the
haploid set (23 chromosomes) & termed as
euploid (Greek, eu = good & ploid = set)
E.g. Haploid gametes & diploid somatic
cells
B. Aneuploidy
Cells that do not contain a multiple of 23
chromosomes are termed aneuploid.
missed or additional chromosomes is
usually one but it is possible more than
one.
Causes: Non-disjunction
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Types of Aneuploidy
1. Monosomy:
Presence of only one copy of
chromosomes in diploid cells
E.g. Turner syndrome
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Types of aneuploidy
2. Trisomy:
The presence of three copies of
homologous chromosomes.
Less sever than mosomies because
body can tolerate excess genetic
materials than deficit
E.g. trisomy 21, trisomy 18, trisomy 13,
klinefelter syndrome.
Autosomal aneuploidy
It is the most clinically important of the
chromosomes abnormalities
Cause: Non-disjunction:
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1. Trisomy 21
Down syndrome (described in 1866 by
John Langdon Down)
1 /700 children affected & compatible
with survival to term
It is due to an extra copy of chromosome
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correlated with age of mother
karyotyping, 47, xx, + 21 or 47, xy, +21
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2. Trisomy 18
Edward syndrome
second most common abnormalities
Occurs 1/6,000 birth
karyotyping, 47, xy, +18 or 47, xx, +18
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3. Trisomy 13
Patau syndrome
Kayotyping 47,xy, +13 or 47, xx, +13
1/500 – 10,000;
Rarely survive more than a year.
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Sex chromosome aneuploidy
1. Monosomy of the X chromosome (X
females)
occur 1/400 males & 1/650 female live birth
infants with some form of sex chromosome
aneuploidy.
Associated with a single X chromosome (45, x)
Turner’s syndrome: Described by Henry Turner in
1938
Phenotypically female but sex organs do not
mature & are sterile
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o Cause of X females:
Deletion of some or the entire short arm
of X chromosome (10 – 20 %)
Absence of paternally derived x
chromosomes i.e. the offspring receives
an x chromosome only from her mother
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2. Trisomy X: XXX females
47, xxx
1/1000 live births;
Suffer from sterility & menstrual
irregularity
Cause: non-disjunction in the mother
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3. Klinefelters syndrome: XXY males
Described by Harry Klinefelter in 1942
Associated with 47, xxy karyotype & 48, xxxy
& 49, xxxxy have also be recorded.
1/2000-1,000 live births;
have male sex organs, but are abnormally
small;
breast enlargement & other female
characteristics; breast dev’t is seen in 1/3rd of
the affected males & leads to an increased
risk of breast cancer
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4. 47, xyy syndrome: XYY males
Karyotye is 47, xyy
taller than average
Involves a slight degree of IQ reduction but
few physical problems
Cause: Non-disjunction in the father
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C. Polyploidy: extra chromosome sets
Presence of a complete set of extra
chromosomes
Include:
triploidy &
tetraploidy
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a. Triploidy
Cells contain 69 chromosomes (karyotype
69, xxx).
Additional chromosomes encode surplus of
gene product, causing multiple anomalies
such as heart & CNS defect
1/10,000 & accounted for 15 % 0f
chromosomal abnormalities at conception.
Thus triploidy conceptions are
spontaneously aborted & those that survived
to term die shortly after birth.
Triploidy contd.
Causes:
Dispermy fertilization of an egg
Fusion of an ovum & a polar body,
Meiotic failure: in which a diploid sperm or
egg cell is produced.
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b. Tetraploidy
92 chromosomes in each cell nucleus.
Karyotype, 92, xxxx
It is rarer than triploidy both at conception &
among live births.
It has been recorded in only a few live births &
those infants survived for only a short period
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Tetraploidy contd.
Causes:
Mitotic failure in the early embryo in which
all of the duplicated chromosomes migrate
to one of the two daughter cells.
It can also result from the fusion of two
diploid zygotes
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2.2 Chromosome structure abnormality
chromosome numbers may be normal, but
abnormalities in chromosome structure can
cause disorders.
Chromosome breakage can lead to a variety of
rearrangements
Include: Deletions, Duplications, Inversions,
translocations
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Deletions
deficiencies or loss of a segment of a
chromosome
Terminal deletion: loss of the end of a
chromosome
Internal deletion: loss of an internal
segment of a chromosome
Effects of deletions
Large deletions are often lethal in the
heterozygous condition (dominant lethal)
Small deletions are often lethal in the
34 homozygous condition recessive lethal)
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Duplications
segment of a chromosome is present in more
than one copy
Duplications usually do not have lethal effects
These mutations occur by recombination
between duplicated regions which increase
the copy number of this segment
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Inversions
a segment of a chromosome is present in
reverse order
Paracentric inversion, both breakpoints are
in the same chromosome arm & the
segment between the breakpoints is
inverted
Per centric inversion, one breakpoint
occurs in each arm & the segment between
the breakpoints is inverted
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Translocations
Pairing of non- homologous
chromosomes in prophase I of meiosis
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Insertion
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Trisomies, non-disjunction & meternal age
Nearly all autosomal trisomies increase with
maternal age as a result of non disjunction.
Little evidence for paternal age effect on non-
disjunction
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Trisomies, non-disjunction & meternal
age cont’d
Mothers
less than 30 ----- risk is < 1/1000
At age of 35 ------ risk increase to 1/400
At age of 40 ------ risk increase to 1/100
At age of 45 ------ risk increase to 1/25
Summary of chromosome aberrations
Quantitative changes (duplications &
deficiencies)
Qualitative changes (inversion,
translocations, ring chromosomes)
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Causes of mutation
1. Spontaneous mutations
Occur without exposure to any obvious
mutagenic agent at the time of DNA
replication.
Mistakes occurring during DNA replication
Occurs 1/10,000 to 1/10 trillion cell
divisions, because enzymes correct
mistakes & damaged DNA
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Causes of mutation cont’d
It may be due to chemical instability of
purine & pyrimidine bases such as
breakage of bonds between sugar &
bases, deamination of cytosine to form
uracil & Mistakes occurring during DNA
replication
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2. Induced mutations
Occur due to mutation causing agents
called mutagens ( chemicals, UV, etc.)
Intentional mutations to study the
expression of the gene
Spontaneous mutation is increased by
exposing an organism to agents that affect a
DNA
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Mutagens
Mutagen is a natural or human-made
agent (physical or chemical) which can
alter the structure or sequence of DNA Or
agents that increase the rate of mutation
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Types of mutagens
1. Chemical
2. Physical (radiation)
3. Biological
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Chemical mutagen
First reported as mutagen in 1942 by
Charlotte Auerbach, who showed that
component of poisonous gas used in
World Wars I & II could cause mutations in
cells. Since that time, many other
mutagenic chemicals have been identified
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Mechanism of Chemical mutagens.
They change the sequence of bases in a
DNA gene in a number of ways;
Mimic the correct nucleotide bases in a
DNA molecule
Remove parts of the nucleotide
Add hydrocarbon groups to various
nucleotides
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Types of chemical mutagens
Include:
1. Base analogue
2. DNA modifying enzymes
3. Intercalating agents
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Base analogs
Compounds that resemble the A, G, C, T
or U
These chemicals may be incorporated
into DNA in place of the normal bases
during DNA replication:
Include: Bromouracil (BU), Aminopurine ,
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Bromouracil (BU):
o Artificial compound extensively used in
research.
o It resembles thymine (has Br atom
instead of methyl group) & will be
incorporated into DNA & occasionally
cytosine (bond shift)
o BU then pair with Adenine like Thymine.
o Overall effect: A:T ---- G:C
Aminopurine
o adenine analog which can pair with T or
(less well) with C;
o Over all effect: A:T to G:C or G:C to A:T
transitions
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DNA modifying enzymes
Chemical which alter structure & pairing
properties of bases by removing or addition
of functional groups on the bases.
e.g. Nitrous acid
It converts Cytosine to Uracil by
deamination (removal of amino group)
Over all effect: G : C ------ to------- A : T
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Intercalating agents:
include: alcidine orange & ethidium
bromide (used in labs as dyes and
mutagens)
All are multiple ring molecules which
interact with bases of DNA & insert between
them.
This insertion causes a "stretching" of the
DNA duplex & the DNA polymerase is
"fooled" into inserting an extra base
opposite to an intercalated molecule.
The result is that intercalating agents cause
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frame shifts.
Physical agents :
Electromagnetic Radiation
Radiation was discovered in 1890's & its effects
on genes were first reported in the 1920's.
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Mechanism of action
1. Ionizing radiation: X-rays
Highly energetic radiation which is
absorbed by the atoms in water
molecules surrounding the DNA. This
energy can eject electrons away from the
atom, forming electrically charged ions
This free radicals, which are highly
reactive molecule that promote chemical
rxn which damages DNA bases.
Radiation can also cause double strand
breaks in the DNA molecule, which the
cell's repair mechanisms cannot put right.
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Mechanism contd.
2. None-ionizing: Uv – light
Sunlight contains Uv radiation which,
when absorbed by the DNA causes a cross
link to form between certain adjacent
bases.
It is less energetic & non-ionizing, but it can
be absorbed by bases of DNA so it can
move electrons from inner to outer orbits
within atoms.
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None-ionizing: Uv – light cont’d
This produces dimers between adjacent
thymine bases in one strand.
These dimer formation block transcription &
DNA replication
lethal if unrepaired b/c it can stimulate
mutation
3. Particle radiation
This term ionization also includes streams
of atomic & subatomic particles emitted by
radioactive elements
These are alpha- & beta-particles
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3. Biological: Transposons
They are mobile genetic elements in bacteria
called jumping genes
They insert them selves into a new regions of
the DNA & cause mutation.
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Consequences of mutation
Most often mutations are lethal
Occasionally a new trait is the result =
source of genetic variation
Impacts treatment on TB & HIV.
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Mechanisms of DNA Repair
• Photoreactivation
• Direct rejoining
• Excision Repair (nucleotide excision repair,
base excision repair)
• Methyltransferase
• Direct insertion
• Damaged DNA replication
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DNA Repair Mechanisms: Photoreactivation
• Enzymes bind specifically to pyrimidine
dimers that form at TT sites in the DNA in
response to exposure to UVC
• Enzyme catalyzes photolysis of
cyclobutane ring linking the adjacent
thymines. The enzyme separates from the
repaired DNA following destruction of the
cyclobutane ring
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DNA Repair Mechanisms: Direct Rejoining
• DNA repair mechanism that functions to
repair single-stranded breaks in the DNA
where strands of DNA are joined directly
through the action of polynucleotide ligase
• The bases on either side of the break
cannot be damaged for this form of repair to
occur
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DNA Repair Mechanisms: Nucleotide Excision
Repair
• Faulty bases are excised from the DNA strand,
attacks bulky DNA adducts & apurinic/apyrimidinic
sites
Mechanism
• Endonuclease enzyme detects the DNA lesion and
makes a single-strand incision
• Exonuclease enzyme excises the defective
segment of DN
• DNA polymerase synthesizes new DNA to fill the
gap
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DNA Repair Mechanisms: Base Excision Repair
• Targets alkylated and deaminated bases in the DNA
and repairs through removal of the damaged base
Mechanism
– DNA glycosylase catalyzes hydrolysis of
glycosylic bond
– AP endonuclease hydrolyzes phosphodiester
bond
– Exonuclease excises deoxyribose-phosphate
residue
– Polymerase and ligase enzymes fill gap in DNA
strand
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DNA Repair Mechanisms: Base Excision Repair
Base excision repair for pyrimidine dimers involves a
number of enzymatic activities:
– Pyrimidine dimer-specific DNA glycosylase
– V-specific endonuclease
– Apyrimidinic (AP) endonuclease
– DNA polymerase, and ligase
– DNA ligase
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DNA Repair Mechanisms: Methyltransferase
• Methyltransferase enzyme functions in the
repair of O6-alkylguanine
• Transfers a methyl group from guanine to
itself, resulting in the inactivation of the
enzyme
• Different tissues express varying levels of
this enzyme, producing different levels of
protection from this form of DNA lesion
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DNA Repair Mechanisms: Direct Insertion
• Accomplished through the action of the
purine insertase enzyme
• Inserts purine into apurinic sites in the
damaged DNA from available free purines
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DNA Repair Mechanisms: Damaged DNA
Replication
• Postreplication repair—Replication
machinery bypasses damaged areas;
gaps containing damaged sites are
subsequently repaired
• Continuous DNA synthesis—Process
generates no gaps as DNA damage is
repaired during synthesis of new DNA.
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Pathologic Manifestations of Defective DNA
Repair
• Various genetic diseases, carcinogenesis,
cell death, and age-related deficiencies
increase susceptibility to DNA repair
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Human Diseases Related to DNA Repair
Deficiency
• Ataxia telangiectasia: (AT) Patients are
sensitive to gamma irradiation; patients develop
neurological & skin lesions
• Fanconi’s anemia: Patients demonstrate
aplastic anemia, growth retardation, &congenital
anomalies; related to a deficiency in repair of
DNA cross-links
• Xeroderma pigmentosum: (XP)
Hypersensitivity to UV light; often develop
04/16/21
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malignancies of the skin
Summary
These processes are regulated & subject to
repair systems that insure the correctness of
information
Mutations are the cause of many disease &
the source of new genetic diversity that
permits adaptation to change
Various agents which cause DNA damage
include: chemical & physical agents such as;
X-rays, ultraviolet light, alkylating agents,
hydrogen ions, chemical mutagens, &
products of cellular metabolism
Summary cont’d
Various types of DNA damage are types
categorized as: pyrimidine dimer, single-or
double-strand breaks, base loss, base
modifications, intercalation and interstrand
cross-links
Mechanisms of DNA repair include:
photoreactivation, direct rejoining, excision
repair (nucleotide and base)
methyltransferase, direct insertion, and DNA
replication
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