Mutations

Mutations can cause cancer.

What Are Mutations?

• Changes in the nucleotide sequence of DNA

• May occur in somatic cells (aren’t passed to
offspring)
• May occur in gametes (eggs & sperm) and be
passed to offspring
 Gene Mutations
• Change in the nucleotide sequence of a gene.
• May only involve a single nucleotide.
• May be due to copying errors, chemicals, viruses,
etc.
Point Mutation
• Change of a single nucleotide.
• Includes the deletion, insertion, or
substitution of ONE nucleotide in a gene.
• If one purine [A or G] or pyrimidine [C or T] is
replaced by the other, the substitution is called a
transition. If a purine is replaced by a pyrimidine
or vice-versa, the substitution is called a
transversion.
 Frameshift Mutation
• Inserting or deleting one or more nucleotides

changes the “reading frame” like changing a sentence.

• Proteins built incorrectly.

• Original:

– The fat cat ate the wee rat.

• Frame Shift (“a” added):

– The fat caa tet hew eer at.

Amino Acid Sequence Changed
 Missense mutations
The new nucleotide alters the codon so as to
produce an altered amino acid in the protein
product.
e.g sickle-cell disease.The replacement of A by T at
the 17th nucleotide of the gene for the beta chain of
hemoglobin changes the codon GAG (for Glu) to
GTG (for Val).
 Nonsense mutations

• The new nucleotide changes a codon that specified

an a. a. to one of the STOP codons (TAA, TAG, or
TGA). Therefore, translation of the messenger RNA
transcribed from this mutant gene will stop
prematurely.

• The earlier in the gene that this occurs, the more

truncated the protein product and the more likely
that it will be unable to function.
 Silent mutations

• Most amino acids are encoded by several different

codons. E.g if the 3rd in the TCT codon for Ser is
changed to any one of the other three bases, Ser will
still be encoded.

• Such mutations are said to be silent because they

cause no change in their product & cannot be
detected without sequencing the gene.
 Suppressor
mutation
• It is a second mutation that alleviates or reverts the
phenotypic effects of an already existing mutation.
• Genetic suppression therefore restores the
phenotype seen prior to the original background
mutation.
Intragenic Suppression

• It results from suppressor mutations that occur in

the same gene as the original mutation.
Base substitution mutations:

Same codon / same nucleotide:

UGG -> UGA -> UGC
trp stop cys

Same codon / different nucleotide:

UGG -> UGA -> CGA
trp stop arg

Different codon in the same gene:

CGA GAG CAU -> CUA GAG CAU -> CUA GAG CCU
arg glu his leu glu his leu glu leu
(wild-type) (1° mutant) (pseudo-revertant)
Frameshift mutations:

5' AUG UGG GGA CCC AAG GGU AGC CCC ... 3' (wild-type)
met trp gly pro lys gly ser pro ...

5' AUG UGG GGG ACC CAA GGG UAG CCC C.. 3' (1° mutant)
met trp gly thr gln gly stop

5' AUG UGG GGG ACC AAG GGU AGC CCC ... 3' (pseudo-revertant)
met trp gly thr lys gly ser pro ...
Intergenic Suppression
• Also known as extragenic suppression relieves the
effects of a mutation in one gene by a mutation
somewhere else within the genome.
• The second mutation is not on the same gene as
the original mutation.
• E.g a mutation which converts a codon specifying
an a.a into a termination codon results in formation
of incomplete polypeptide which are usually inactive.
A mutation in another gene can occur which codes
for a tRNA that reads a termination codon. So
another a.a gets inserted & no chain termination
occurs & enzyme got may be active.
Cell death and its
regulation
 Homeostasis

• Constant number of cells in an organism.

• Cell death = Cell proliferation

Cell Growth
Survival
Death Proliferation

Cell death must be balanced by cell renewal & most

tissues have stem cells that replace cells that have
been lost.
Apoptosis (?)
Programmed cell death.
Disintegration of cells into membrane-bound
particles that are then eliminated by phagocytosis
or shedding.
Occurs following irreparable damage to the cell or
when “told” to do so.
Responsible for maintaining constant cell no.
Provides a defense mechanism by which damaged
& potentially dangerous cells are eliminated e.g
virus infected cells, cells with damaged DNA
(mutations) which may be harmful i.e cause
cancer.
 Examples of apoptosis

Embryonic Development
– Tadpole tail
– Digit sculpting

Adult
– Bone marrow cells
– Intestinal cells
– Menstruation
 Events of Apoptosis

Necrosis Accidental death due to acute injury

PCD Proceeds by a series of cellular changes

called apoptosis

1. Chromosomal DNA fragmented

2. Chromatin condenses
3. Nucleus breaks into small pieces
4. Cell breaks into membrane-enclosed fragments (apoptic
bodies)
Apoptic cells & cell fragments are recognized &
phagocytosed by both macrophages & neighbouring
cells. They have so called “eat me” signals on the cell
surface e.g Phosphatidylserine on cell surface.
 Caspases
- Ultimate executioners of PCD.
- Cleave more than 100 diff. proteins e.g an inhibitor of a
DNase, nuclear lamins, cytoskeleton proteins, golgi matrix
proteins, etc.
- All are synthesized as inactive precursors that are
converted active by proteolytic cleavage by other caspases.
- Initiator caspases are activated directly in response to
various signals of apotosis. They than cleave &activate the
effector caspases which digest target proteins.
- Caspase-9 is activated as a complex with Apaf-1 and
cytochrome c in the apoptosome. It than cleaves & activates
effector caspases e.g caspase-3 & caspase-7, eventually
leading to cell death.
Mechanism of apoptosis: caspases
 Regulation of apoptosis

Mammals have ~20 proteins related to Bcl-2 which are

divided into 3 gps:
1. Antiapoptotic family members. E.g Bcl-2 which inhibit
apoptosis. They have 4 Bcl-2 homology domains (BH1 –
BH4).
2. Proapoptotic multidomain members. E.g Bax & Bak.
Have 3 BH domains.
3. Proapoptotic BH3-only members. E.g Bid, Bad, Noxa,
Puma, Bim have only the BH3 domain.
In normal cells, the BH3-only proapoptotic proteins are
inactive & the multidomain proapoptotic proteins are
inhibited by interaction with antiapoptotic proteins.

Cell death signals

Activate BH3-only proteins

Interact with antiapoptotic proteins

Activate the multidomain proapoptotic proteins

Cell death

Bax & Bak oligomers in OM of Mitochondria

Release of Cyt. C

Caspase activation
Bcl-2 proteins regulate caspase activation
 Inhibitor of apotosis (IAP)

Interact with caspases

Inhibit them or target them for ubiquitin mediated

proteolysis

Suppress apoptosis

A variety of signaling pathways

Regulate apoptosis by controlling expression or activity of

proapoptotic members
e.g DNA damage: induce p53
growth factor : activation of PI3-kinase/Akt signaling
polypeptides: activate death receptors PCD
 Alternative pathways of PCD

-Autophagy

-Regulated necrosis