MUTATIONS

Friday, 9 December 2022 9:57 pm

• are permanent modifications in the base sequence of DNA

• If germ cells are affected transmitted to the progeny
• In somatic cells not transferred to progeny but important in cancer causation and
congenital diseases

Germline mutation - change occurs during the DNA replication that precedes meiosis.
Somatic mutation - happens during DNA replication before mitotic cell division

Different types of mutation

• Point mutation
• Frameshift mutations
• Trinucleotide repeat mutations

POINT MUTATION
• occur at the level of one or a few bases of DNA

May involve either:

• Substitution
○ Transition vs transversion
• Deletion
• Insertion

MUTATIONS
• At the level of a gene, mutations involve dozens to thousands of bases
• At the genomic level mutations include: deletions or duplications of hundreds of
thousands to millions of bases, up to chromosome rearrangements and aneuploidies

SUBSTITUTION
• Transition - substitution of a purine by another purine or of a pyrimidine by another
pyrimidine
• Transversions - exchanges of purines and pyrimidines

COPY NUMBER VARIATION (CNV)

• involves large deletions and insertions of various lengths created by unequal crossing
over between misaligned segments of repetitious DNA or by non-homologous end
joining
• Unequal crossing over is the origin of X linked anomalous colour vision
• Activation of enhancers and silencers can cause phenotypic variation in expression of
the genes they control
• involves amplification or deletion of a large segment of DNA
• It is associated with many pathologies and sometimes distinguishes the genomes of MZ
twins

DYNAMIC MUTATIONS
• involve expansion of triplet repeat sequences and can undergo further expansion or
contraction from generation to generation.

SUBSTITUTION
• involves replacement of a base pair
• silent mutation/substitution
○ If the amino acid encoded by the new codon is the same
○ Mutations that do not change amino acid sequences:
• missense mutation
If a different, amino acid is encoded

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 ○ If a different, amino acid is encoded
○ BP substitution producing and change in a single AA

Conservative mutation Deleterious mutation

• missense mutations that do not
alter the chemical properties of the
protein
• In some cases, though, heterozygosity of a
deleterious mutation may create selective
advantage
• A notable example is the substitution of
the sixth codon in the β globin chain
responsible for sickle cell anemia , which in
heterozygotes confers resistance to
malaria

• nonsense (‘non sense’) mutation or premature termination

○ Substitution can create a STOP codon, causing translation to come to a
premature halt
○ BP substitution producing a stop codon

FRAMESHIFT MUTATION
• If a deleted or inserted segment is of other than a multiple of three bases , the
translational reading frame is disrupted
• This causes the protein produced to be entirely erroneous ‘downstream’ (3 ′′) of the
deletion
• The most common mutation causing Tay Sachs disease is a four base insertion causing
a frameshift and leading to premature termination , so that no functional
hexosaminidase A is synthesized

DUPLICATIONS
• Duplications of whole genes can lead to disease
• Charcot Marie Tooth disease
○ a peripheral nervous system disease involving progressive degeneration of
peripheral nerves, leading to atrophy of distal limb muscles
○ It can be caused by a variety of types of mutation, but about 70% have a large
duplication of chromosome 17 that includes the gene PMP22, encoding a
peripheral myelin protein.
○ This contributes to demyelination, whereas deletion of the same region causes a
paralytic response to pressure (hereditary neuropathy with predisposition to
prpressurealsies)

TRINUCLEOTIDE REPEAT MUTATIONS

• Set of genetic disorder caused by trinucleotide repeat in certain gene’s exceeding
normal, stable threshold
○ Fragile X syndrome
○ Huntington's disease
○ Myotonic dystrophy

CAUSES OF MUTATION
Chemical causes Physical causes
• Nitrous acid • Radiation
• Alkylating agent • Xray & Ultraviolet light
• 5 bromouracil • Certain viruses
• Antiviral drug iododeoxyuridine
• Benzopyrene in tobacco smoke

ENVIRONMENTAL MUTAGENS PRINCIPAL MEANS OF EXPOSURE

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ENVIRONMENTAL MUTAGENS PRINCIPAL MEANS OF EXPOSURE
• Smoke • Inhalation
• Paints • skin absorption
• Petrochemicals • ingestion
• Pesticides
• Dyes
• Foodstuffs
• Drugs

PROMUTAGENS
• Nitrates and nitrites
• Are converted into mutagens by body chemistry

CHEMICAL MUTAGENS
• Base analogues
○ 2 amino purine is incorporated into DNA in place of adenine, but pairs as
cytosine, causing a substitution of thymine by guanine in the partner strand
○ 5 bromouracil (5 BU) is incorporated as thymine, but can undergo a tautomeric
shift to resemble cytosine, resulting in a transition from T-A to C-G
• Chemical modifiers
○ Nitrous acid converts cytosine to uracil, and adenine to hypoxanthine, a
precursor of guanine
○ Alkylating agents modify bases by donating alkyl groups
• Intercalating agents
○ The antiseptic proflavine and the acridine dyes become inserted between
adjacent base pairs, producing distortions in the DNA that lead to deletions and
additions
• Others
○ Cytosine adjacent to guanine (‘CpG’) is prone to methylation by
methyltransferase enzymes to 5 methylcytosine which is unstable and prone to
deamination to thymine.
○ CpG pairs tend to undergo transition to
TpA , despite scrutiny by the DNA repair enzymes

ELECTROMAGNETIC RADIATION
• Particulate discharge from radioactive decay:
○ alpha particles (helium)
○ beta particles (electrons)
○ gamma rays
• mutagenicity of subatomic particles depends on their speed, mass and electric charge
• energy & mutagenicity of electromagnetic radiation increases w/ decreasing
wavelength
• All radiation beyond UV causes ionization by knocking electrons out of their orbits

ULTRAVIOLET LIGHT
• non visible, short wavelength fraction of sunlight responsible for tanning
• exerts a mutagenic effect by causing dimerization (linking) of adjacent pyrimidine
residues, mainly T-T (but also T-C and C-C)
• does not cause germline mutations
• Major cause of skin cancer
• homozygotes for the red hair allele
○ white, freckled skin.
○ phaeomelanin pigment provides a poor sun screen and releases free radicals on
UV exposure
• Most UV radiation from the sun is blocked by a layer of ozone in the upper
atmosphere, currently in danger of destruction by industrial fluorocarbons

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 atmosphere, currently in danger of destruction by industrial fluorocarbons

ATOMIC RADIATION
• Natural background radiation
○ This varies with local geology
○ The most abundant radioisotopes are Potassium 40 and Radon 222 gas
○ Radon contributes 55% of all natural background radiation and may be
responsible for 2500 deaths per annum in the UK
• Cosmic rays
○ a major theoretical hazard for aircrews
○ intensity increases with altitude
○ Exposure during a return flight between England and Spain is said to equal five
chest X rays
• Man-made radiation
○ fallout from nuclear testing and power stations
○ Radiation workers are at particular risk
○ periosteum seeking isotopes present a major risk of leukemia
• X-rays
○ account for 60% of man made and 11% of total radiation exposure
○ mutagenize DNA either directly by ionizing impact, or indirectly by creating
highly reactive free radicals that impinge on the DNA
○ These can be carried in the bloodstream and harm cells not directly exposed.

BIOLOGICAL EFFECTS OF RADIATION

• Electromagnetic radiation
○ damages proteins and above 1 gray kills cells
○ One gray( Gy ) is the amount of radiation that causes 1 kg of tissue to absorb 1
joule of energy
○ At the chromosomal level it causes major deletions, translocations and
aneuploidy
○ It causes single and double strand breaks in DNA and base pair destruction
• X rays damage chromosomes most readily when they are condensed, which is why
they are most harmful to dividing cells, including the progenitors of sperm
• The offspring of older men have a many fold risk of genetic disease, as the DNA in their
sperm has been copied many times
• The high testicular temperature caused by clothing (6C above unclothed) also
contributes to the present mutation rate in human sperm
• the first 7 days of life the embryo is ultrasensitive to the
• mutagenic and lethal effects of X rays
• over weeks 2-7 teratogenic effects come to the fore
• Childhood leukemia can be induced by exposure at gestational weeks 8 to 40
• At 1 Gy , X rays cause a 50% reduction in WBC count, while whole body irradiation of
4.5 Gy kills 50% of people
• Therapeutic doses of up to 10 Gy are used against cancer cells
• Radiation damage is cumulative and there is no lower baseline off effect

SAFETY MEASURES WHEN USING X RAYS

• Previous X ray exposure should be reviewed before requesting further X rays
• 28-day rule
○ a woman of child bearing age should be X rayed only if she has had a period
within the last 28 days
• 10-day rule
○ for high radiation dose procedures such as abdominal or pelvic CAT scan and
barium enema , the examination should be postponed to the first 10 days of the
menstrual cycle

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CLASSIFICATION OF GENETIC DISEASES
1. Single gene defects / Mendelian disorders
2. Disorders with multifactorial or polygenic inheritance
3. Cytogenetic disorders
4. Disorders showing atypical patterns of inheritance

MENDELIAN DISORDERS
• A genetic disease caused by a single mutation on the DNA structure
• Causes a single basic defect with pathologic consequences
• Patterns of inheritance:
○ Autosomal dominant
○ Autosomal recessive
○ X linked recessive
○ X linked dominant

AUTOSOMAL DOMINANT DISORDERS

• Manifested in heterozygous states
• Individuals with these diseases usually have one affected parent
• Variable to late onset
• Usually involve non enzymatic proteins
○ Proteins involved in metabolic pathway regulation
○ Structural proteins

RULES FOR AUTOSOMAL DOMINANT INHERITANCE

• (1) Both males and females express the allele and can transmit it equally to sons and
daughters
• (2) Every affected person has an affected parent (‘vertical’ pattern of expression in the
pedigree). Direct transmission through three generations is practically diagnostic of a
dominant.
• (3) In affected families, the ratio of affected to unaffected children is almost always 1 :
1
• (4) If both parents are unaffected, all the children are unaffected

AUTOSOMAL DOMINANT INHERITANCE

• vertical transmission of the disease phenotype
• NO skipped generations
• equal numbers of affected males and females
• Father-to-son transmission may be observed

MARFAN SYNDROME
• Mutation in the fibrillin gene
• excessive elasticity of fibrillin 1

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 • excessive elasticity of fibrillin 1
• Fibrillin is important component in microfibrils
• Affects the skeleton, eyes and CVS
• Aortic root aneurysm & ectopia lentis

• Tall stature
• Long fingers
• Pigeon breast deformity
• Hyperextensible joints
• High arched palate
• Bilateral subluxation of lens
• Floppy mitral valves
• Aortic aneurysm & dissection

EHLER-DANLOS SYNDROME
• Cutis hyperelastica
• Defects in collagen synthesis
• Fragile, hyperextensible skin
• Hypermobile joints
• Grotesque contortions
• Rupture of internal organs (colon, cornea & large arteries)
• Poor wound healing

FAMILIAL HYPERCHOLESTEROLEMIA
• One of the most common mendelian disorders
• Mutation on the LDL receptor gene
• Impaired LDL transport into cells
• Increased risk of atherosclerosis & coronary artery disease
• Xanthoma formation

Autosomal recessive disorders

• Largest group of mendelian disorders
• Affected individuals usually have unaffected ( carrier ) parents
• Uniform, early age of onset
• Usually involve enzymatic proteins

RULES FOR AUTOSOMAL RECESSIVE INHERITANCE

• (1) Both males and females are affected
• (2) There are breaks in the pedigree and typically the pattern of expression is
‘horizontal’ (i.e. sibs are affected but parents are not)

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 ‘horizontal’ (i.e. sibs are affected but parents are not)
• (3) Affected children can be born to normal parents, usually in the approximate ratio of
one affected to three unaffected
• (4) When both parents are affected all the children are affected, unless mimic genes
are involved
• (5) Affected individuals with normal partners usually have only normal children.

AUTOSOMAL RECESSIVE DISORDERS

1. Cystic fibrosis
○ Autosomal homozygous recessive
○ Patient has to inherit a recessive gene from each parent
○ Both parents are carriers (heterozygous)
○ 25% chance in siblings
○ 1/~2,500 Caucasians;
○ 1/15,000 African Americans
○ 1/30,000 Asian Americans
○ AR, 7q31
○ Phe508del accounts for 70%
○ Newborns 10 20% have a thick plug that blocks the colon called meconium ileus
○ pancreatic insufficiency intestinal malabsorption, anemia and failure to thrive,
rectal prolapse and blockage of liver ducts
○ sweat is very salty
○ Almost all males have congenital bilateral absence of the vas deferens
○ chronic obstructive airway disease due to thick mucus
○ basic defect: cystic fibrosis transmembrane conductance regulator (CFTR)
protein responsible for controlled passage of chloride ions through cell
membranes

2. Hemochromatosis
3. Glycogen storage disease
4. Sickle cell anemia
5. Thalassemia
6. X linked recessive disorders

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 6. X linked recessive disorders

PHENYLKETONURIA
• AR homozygous
• PKU homozygotes are fair haired with blue eyes.
• Children have convulsions and become severely intellectually impaired ,
phenylalanine (PA) accumulates in the blood and related metabolites are excreted in
the urine
• The basic cause is deficiency in phenylalanine hydroxylase (PAH) necessary for
conversion of PA into tyrosine
• mousy’ smell due to phenylacetic acid in the sweat and urine, and muscular
hypertonicity

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