EXAM2_L15_DNA_REPAIR

Question 1

Mutations in germ (sex) cells result in?

Mutation in somatic cells result in?

Answer 1

germ cell mutations - inherited diseases

somatic cell mutations- Cancer

Question 2

How do mutations become a permanent part of the genome?

Answer 2

ie- methylation of guanine would change its hydrogen bonding to bind with T better than C.

-Once DNA replicates the daughter strands its permanent

Question 3

Single base substitution-

Answer 3

Ie sickle cell anemia
GAG=Valine normal
GTG= Glutamine sickle cell

Question 4

DNA pol Proofreading- reduces errors to 1 in 10^7

What used if missed?

Answer 4

• DNApol cleaves wrong nucleotide before adding another

- if missed; Strand-directed Mismatch Repair used (MMR)

Question 5

MMR 4 steps. when used?

Answer 5

detects distortions in DNA helix from errors missed by proofreading-occurs during replication
1. MSH protein binds the buldge in dsDNA
2. MLH binds to MSH and finds closest nick
3. MLH triggers degradation from error to nick
4. gap filled by DNApol & sealed by DNAligase
MSH -SITS MLH-LOOKS

Question 6

HNPCC- What -where-how

Answer 6

Hereditary non-polyposis Colon Cancer

• tendency to develop cancer is inherited
• (occurs without polyps present)
• due to mutation in Msh1 (60%) and Mlh1 (30%) genes
• b/c dna repair is broken adenoma–>carcinoma in only 2-3 years vs usual 8-10years seen in non-HNPCC patients

Question 7

What 3 ways for SS break repair?

What 2 ways for DS break repair?

Answer 7

SS break repair:
1. Site-directed MMR
2. Base Excision Repair (BER)
3. Nucleotide excision repair (NER)
DS break repair
1. Non-homologous end joining (NHEJ)
2. Homologous Recombination (HR)

Question 8

BER

Answer 8

• DNA glycosylases remove all unnatural bases created by spontaneous deamination
• leison specific (~8 types in humans)
• deamination cytosine to uracil 100 bases per cell/day

Question 9

BER 3 steps

Answer 9

1. Glycosylase cleaves base leaving abasic sugar
2. AP endonuclease removes sugar-phosphate backbone
3. DNA pol adds new nucleotides DNA ligase seals nick

Question 10

NER 4 steps.

ie- pyrimidine dimers– 2 adjacent thymine or cytosine residues link together covalently

Answer 10

1. Multi-enzyme complex find distortion of helix
2. nuclease cuts backbone on both sides of distortion
3. helicase separates damage oligonucleotide
4. DNApol uses undamaged strand as template

Question 11

NHEJ- function- problems

Answer 11

Each strand uses each other as template for DNA replication

• DNA ligase seals the gap
• prone to mutagenic and error b/c it removes several base pairs at the break site

Question 12

Homologous Recombination HR

Answer 12

Uses sister chromatid or homologous chromosome as a template
-ds breaks during cell division (not replication) when duplicated chromosomes haven’t yet separated.
-best for post replication repair
ERROR FREE

Question 13

Familial breast cancer- What proteins? Chance of cancer if inherited?

Answer 13

BRCA1 and BRCA2 proteins normally involved in dsDNA break repair

Women are 40-80% more likely to develop breast/ovarian cancer if mutated/deficient BRCA1 and BRCA2 genes are inherited

Question 14

4 Endogenous Sources of DNA damage

Answer 14

1. DNA replication error
2. Deamination
3. Base Hydrolysis
4. Oxidation

Question 15

2 exogenous sources of DNA damage

Answer 15

1. UV damage

2. Carcinogens: DNA adducts

Question 16

Damage by Deamination- what- how repaired

Answer 16

removal of amino group (cytosine>Uracil)
-Uracil DNA glycosylase enzyme scans and removes uracil bases in DNA
-Alters what a base will pair with
_repaired by BER

Question 17

Base Hydrolysis- what - how repaired?

Answer 17

Depurination- removal of a purine base from DNA strand

• glycosidic bond of sugar/base can spontaneously hydrolyze leaving sugar without a base (apurinic site)
• if not repaired>deletion mutation
• BER used begins w. AP endonuclease

Question 18

Oxidation of Base- what is? repair type?

Answer 18

Reactive oxygen species (O2,H2O2,OH*)
-cause mispairing
-complete loss of base from ribose
-induce single and double-strand breaks
use BER

Question 19

UV irradiation damage- What is? Repair type?

Answer 19

UV can induce pyrimidine dimer formation
-adjacent thymine bases can link forming cyclobutane ring
Uses NER because of double bond between adjacent bases

Question 20

Xeroderma Pigmentosum (XP)- What is?

Answer 20

• Hereditary
• Inability to repair DNA damage from UV b/c defective NER
• XP patients 1000x greater risk of developing skin cancer and 100,000x greater risk of developing carcinoma of tip of the tongue

Question 21

DNA Adducts- Two types and examples of each? Type of repair used?

Answer 21

Chemical carcinogens- natural and synthetic chemicals

• add bulky adducts to DNA
  1. benzopyrene (coffee; coal tar)
  2. Acetaldehyde (tobacco) & occurs naturally when converting alcohol in liver.

-USES NER b/c lesion is too bulky for BER to be effective

Question 22

DIAGRAMS on pg 25 and 26 should be used

Answer 22

Really good!