CHAPTER 14

MUTATION, DNA REPAIR

AND CANCER

cmchen@ym.edu.tw
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Permission required for reproduction or display.

Mutation
A heritable change in the genetic material
Essential to the continuity of life

Source

of variation for natural selection

New mutations are more likely to be harmful

than beneficial
DNA repair systems reverse DNA damage
Cancer is a disease caused by gene mutations

Point mutation examples

Base substitution

5 GGCGCTAGATC 3

5 GGCGCGAGATC 3

3 CCGCGATCTAG 5

3 CCGCGCTCTAG 5

Add or delete a single base pair

5 GGCGCTAGATC 3

5 GGCAGCTAGATC 3

3 CCGCGATCTAG 5

3 CCGTCGATCTAG 5
3

Consequences of Point Mutations Within the

Coding Sequence of a Structural Gene
Mutation in
the DNA

Effecton
polypeptide

None

None

Example*

ATGGCCGGC CCGA AAGAGA CC

Met

Base
Silentcauses
substitution no change

Base
Nonsense
substitution changes to a
stop codon
Addition (or
deletion) of
single base

Frameshift
produces a
different amino
acid sequence

Gly

Pro

Lys

Glu

Thr

ATGGCCGGC CCCAAAGAGA CC
Met

Base
Missense
substitution changes one
amino acid

Ala

Ala

Gly

Pro

Lys

Glu

Thr

ATGCCCGGC CCGA AAGAGA CC

Met

Pro

Gly

Pro

Lys

Glu

Thr

A TG G C C GG C C C GTA A GA G A C C
Met

Ala

Gly

Pro

STOP

ATG G C C GG C A C C GA AA GAG A C C
Met

Ala

Gly

Thr

Glu

Arg

Asp

*DNA sequence in the coding strand. This sequence is the same as the mRNA sequence
except that RNA contains uracil (U) instead of thymine (T).

Gene mutations may affect

amino acid sequences
Silent

mutation
Missense mutation
Nonsense mutation
Frameshift mutation

Sickle-cell anemia is the result of a single

amino acid substitution

Sickle-cell anemia and malaria (

Gene mutations outside of coding

sequences

Promoter
May

Transcriptional response element/ operator site

May

increase or decrease the rate of transcription

alter regulation of transcription

Splice junctions
Mutations

at the boundaries between introns and

exons can prevent proper splicing

Translational response elements

May

prevent proper translational regulation

Intergenic sequence

Germ-line or somatic cell mutations

Spontaneous or induced mutations

Spontaneous mutations result from
abnormalities in biological processes
Rates vary from species to species and from
gene to gene
Expected rate of background mutation
approximately 1 mutation for every 1 million
genes

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Spontaneous or induced mutations

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Examples of Mutagens

12

Base deamination: replacing amino groups with

keto groups

13

Ionizing radiation creates free

radicals
X rays and gamma rays
Base deletions, breaks in 1 or
both DNA strands
Nonionizing radiation has less
energy and can only penetrate the
surface

UV

rays can cause formation of

thymine dimer causing gaps or
incorporation of incorrect bases

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Ames test

Uses Salmonella typhimurium that cannot

synthesize histidine due to a point mutation
Bacteria

cannot grow unless histidine is added to

medium
Or, a second mutation occurs that fixes the
original, allowing synthesis of histidine

Test monitors rate at which second mutation

occurs
Allows

us to test and quantify mutagenicity

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Control

1 Mix together the Salmonella

typhimurium strain, rat liver

extract, and suspected
mutagen and incubate. The
suspected mutagen is omitted
from the control sample. The
rat liver extract is added
because liver enzymes
sometimes convert
chemicals into mutagens.

Plate the mixtures onto petri

plates that lack histidine.
Incubate overnight to allow
bacterial growth.

Rat liver
extract

Rat liver
extract

S. typhimurium
strain (requires
histidine)

S. typhimurium
strain (requires
histidine)

Suspected
mutagen

His(-) plate

A large number of colonies

suggests that the suspected
mutagen causes mutation.

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DNA repair
All living organisms must have the ability
to repair changes that occur in DNA in
order to minimize mutation
Requires
DNA damage be detected
Repair of DNA damage

17

Types of repair

Direct repair
Enzyme

removes a modification (for example

an alkyl group)

Altered DNA strand removed and new

segment synthesized
Nucleotide

excision repair
Methyl-directed mismatch repair
The Nobel Prize in Chemistry 2015 was awarded
jointly to Tomas Lindahl, Paul Modrich and Aziz
Sancar "for mechanistic studies of DNA repair".

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Nucleotide Excision Repair (NER)

Four key proteins

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NER and human genetic disease

NER systems were discovered in
humans by the analysis of
genetic diseases that affect DNA
repair
Photosensitivity is a common
characteristic in all three
syndromes because of an
inability to repair UV-induced
lesions

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Cancer
Cancers originate from a single cell
Mutates so that cell grows abnormally
Tumor- an overgrowth of cells with no
useful purpose
Tumor may begin as a benign or precancerous

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Cancerous stage
Malignant

- lost normal growth regulation

Invasive- can invade healthy tissue
Metastatic- can migrate to other parts of the
body

Left untreated, malignant cells will cause

the death of the organism
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Cancer development is a multistep process

23

The epidermal growth factor signaling

pathway

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Ras releases GDP

and then binds GTP
to become active.

GTP
GDP

Ras

Active Ras
protein

Inactive Ras
protein
GDP

GTP

The active
Ras protein
participates
in a signal
transduction
pathway that
promotes cell
division.

Pi
GTP hydrolysis returns
Ras to an inactive state.

Intracellular

signaling protein that hydrolyses GTP

When GTP is bound, Ras promotes cell division
Oncogenic mutations may decrease ability of Ras to
hydrolyze GTP or exchange GDP/GTP faster
Both keep signaling pathway constantly on
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Cancer-causing genes

Oncogene
When

a mutation causes this gene to be

overactive, uncontrolled cell growth occurs.

Tumor-suppressor gene
Normally,

this gene encodes a protein that

prevents cancer
If a mutation eliminates this function, cancer
can occur
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Proto-oncogene

1.
2.
3.
4.

Normal gene that, if mutated, can

become an oncogene
4 common genetic changes
Missense mutations
Gene amplifications
Chromosomal translocations
Retroviral insertions
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Missense mutations convert a proto-oncogene

to an oncogene
Proto-oncogene

Promoter

Coding sequence

A change in the amino acid sequence of a protooncogene protein may cause it to function in an
abnormal way. For example, missense mutations
can convert ras genes into oncogenes.
Missense mutation

(a) Missense mutation

A missense mutation that

converts the normal ras
gene into an oncogene
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Gene amplifications
Abnormal increase in
copy number results
in too much of the
encoded protein
Many human cancers
are associated with
the amplification of
particular protooncogenes

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Chromosomal
translocations
Two

different chromosomes
break, and the ends of the
broken chromosomes fuse
with each other incorrectly
Very specific types of
chromosomal translocations
have been identified in
certain types of tumors
Chimeric genes are
composed of two gene
fragments fused together
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ReciprocaltranslocationofChr22andChr9inchronic
myelogenousleukemia(CML)

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Retroviral insertions
Viral

DNA may insert into

a host chromosome in
such a way that a viral
promoter and response
elements are next to a
proto-oncogene
Alternatively, a virus may
cause cancer because it
carries an oncogene in
the viral genome

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Tumor-suppressor genes

Role to prevent cancerous growth

Proteins encoded by tumor-suppressor genes
usually have one of two functions
1. Maintenance of genome
2. Negative regulators of cell division

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Cell cycle and its checkpoints

Interphase
M

Phase

Proteins

called cyclins and cyclin-dependent protein

kinases (cdks) are responsible for advancing a cell
through the four phases of the cell cycle

p53 and checkpoint control

3 critical checkpoints in eukaryotes

G1 checkpoint (restriction point)
G2 checkpoint
Metaphase checkpoint

Loss of checkpoint
function can lead to
mutation and cancer
p53 is a G1 checkpoint
protein
Expression of the p53
gene is induced when
DNA is damaged

Rb gene
First tumor-suppressor gene to be
identified in humans by studying patients
with a disease called retinoblastoma
Some people have an inherited form that
occurs early
Other forms caused by environmental
agents occur later in life

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Rb negatively regulates the

transcription factor E2F

CdK

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Two-hit model for retinoblastoma

People have two copies of the Rb gene, one from each

parent

Individuals with the inherited form of the disease have one

mutant gene already from one parent
One additional mutation will cause disease
Tends to occur early in life

People with the noninherited form of the disease must have

two mutations in the same retinal cell to cause the disease
The noninherited form of this disease is expected to
occur only rarely and late in life
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Loss of Tumor Suppressor Genes

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Loss of tumor-suppressor gene function

3 common ways
Mutation

occurs specifically within a tumorsuppressor gene to inactivate its function

Chromosome loss may contribute if the
missing chromosome carries one or more
tumor-suppressor genes
Abnormal methylation of CpG islands near
promoter regions

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Lung Cancer Progression

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Dysplasia
Cilia

Abnormal cells

Columnar
epithelium

Basement
membrane

Basal
cells
Connective
tissue

Connective Basement
tissue
membrane

Dysplasia (initially precancerous, then cancerous)

Normal lung epithelium

Abnormal
cells

Columnar epithelium
Basal cells
Connective
tissue

Hyperplasia

Basement
membrane

Cancerous
invasion

Hyperplasia

Connective
tissue
Invasive cancerous cells that can metastasize

Basal cells

Connective
tissue

Basement
membrane

Invasion and Metastasis

Loss of ciliated cells

(15): Dr. Oscar Auerbach, reproduced with permission.

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