Mechanisms of DNA Repair Flashcards Preview

Exam 1 Information > Mechanisms of DNA Repair > Flashcards

Flashcards in Mechanisms of DNA Repair Deck (30):
1

DNA Damaging Agents

-Radiation damage
-UV damage
-Chemical modifications of bases
-Substances that intercalate between double-strand DNA
Form interstrand crosslinks in DNA helices
-Metabolically activated carcinogens
-Oxidation damage

2

Base modifications

Spontaneous lesions- Will result in mispairing

Bromouracil vs cytosine

3

Bromouracil

-Misincorporated for thymine
-Base pairs with guanine

4

Deamination of cytosine

Looks like uracil. Base pairs with Guanine

5

Deamination or depurification results in:

One mutation and one normal DNA molecule after replicaiton

6

Mitochondrial DNA mutations is ____ efficient than nuclear DNA repair

Less efficient. Can cause:
-Cancer
-Aging
-Degenerative diseases

7

Excision repair

Cannot correct chromosomal aberrations
-Involves cutting out and resynthesizing the area of DNA surrounding the damage.
-Mechanism triggered depends on type of damage to DNA
-Undamaged DNA strand used as template

8

Nucleotide Excision Repair

Repairs the majority of bulky lesions in DNA due to UV-induced poto-products and addition of bulky adducts derived from cisplatin and 4-nitroquinoline oxide
Removes lesions such as pyrimidine dimers, photo-adduct products, alkylated bases

-Example of falilure: Xeroderma Pigmentosum

9

Base excision repair

Single nucleotide is replaced during the repair process

Abnormal bases are removed by specific DNA glycosylases

Removes abnormal bases arising from either a deamination or depurination reaction; adduct formation (e.g. methyladenosine), oxidized bases (e.g. 8-oxodG), saturated and ring-fragmented bases.

10

Mismatch Repair enes

Removes base mismatches, corrects insertion, deletions

-Example of mutation: hereditary nonpolyposis colon cancer

11

Xeroderma Pigmentosum

Defective nucleotide excision repair (excision endonuclease)
-Rare autosomal-recessive inherited disorder
-characterized by extreme skin sensitivity to UV light, abnormal skin pigmenation
-Skin cancers
-Some patients develop neurological symptoms

12

Reactive oxygen species (ROS)

Mitochondrial DNA is prone to oxidative damage, as mitochondria are the site of ROS generating respiratory chain.

13

Cockayne syndrome

Patients with Cockayne syndrome have sun sensitivity, short stature and progressive neurological degeneration, along with early senility. However, the condition is not associated with cancer. Cockayne syndrome is recessively inherited and is due to a defect in transcription-coupled NER. This is a variant of NER and operates during transcription in terminally differentiated cells such as neurons.

14

Steps of NER-- affected area is removed and patched

Recognition of defect by ERCC1, XPA, and XPF
A. Nuclease cleavage of phosphodiester bond and "excision" of several nucleotides adjacent to the dimer
B. "Gap" is filled by DNA polymerase in which the appropriate dNTP is added to the 3'-OH end of the clipped DNA (opposite strand will be provided to correct coding information)
C. Final joining (DNA ligase) of the 3'-OH end of the last base added to close the "gap"

15

Steps of NER-- thymine dimer

A.. Location of Thymine dimer
B. Cut by endonuclease
C. Bases removed by helicase
D. New bases added. Ligase seals the ends

16

Steps of BER-DNA Glycosylase

A. DNA Glycosylase recognizes and removes the damaged base from the DNA backbone to form an abasic site (AP)
B. AP site is recognized and cleaved by AP endonuclease which introduces a DNA strand break
C. DNA Poly B adds the appropriate dNTP to fill the nucleotide gap
D. Nick is sealed by DNA ligase

Note: BER repairs oxidative damage in mitochondria

17

Mitochondrial DNA is repaired by

BER

DNA glycosylase, AP endonuclease, DNA polymerase gamma, DNA ligase

18

Mismatch Repair Genes (MMR)

Corrects misincorporated and deleted/inserted bases, particularly those introduced by strand slippage of DNA polymerase

19

When are mismatched sequences corrected?

After replicatoin-- by MMR. Scans the newly synthesized strand

20

Lynch syndrome

Also known as hereditary nonpolyposis colon cancer

Defect in MMR protein results in susceptibility to cancer of the colon and ovaries

Tumors will display microsatellite instability: High instability of short repeated sequences

21

Direct Repair

Dealkylation of guanines: Guanine alkyltransferases remove methyl/ethyl groups from alkylated guanines.

22

Recombination Repair

When damaged DNA escapes repair, the resultant distortion disrupts the process of replication. During DNA replication, the polymerase skips over the distortion, creating a gap on the newly synthesized strands. An exchange process takes place to remove the gap with a homologous segment from the intact strand. The resultant gap on the “donor” stand is filled by repair synthesis as replication proceeds.

23

Repairing Double Stranded Breaks

Homologous recombination repair (HHR)
Non-homologous end joinings (NHEJ)

24

Homologous recombination repair

may operate during DNA replication where it restarts broken DNA replication forks. A homologous DNA strand is used as a template to copy the missing DNA sequences. However, the requirement for a homologous template for repair is a limitation of double-strand-break repair by homologous recombination.

25

Non-homologous end joinings

Simply ligates the two ends together, hoping no DNA was lost during the process

Uses proteins:
- Ku
-DNA-dependent Protein Kinase
-BRCA 1/2


involves rejoining the two ends of the break without relying on a template sequence. This process is error prone as DNA sequences may be lost prior to the repair. NHEJ is also associated with chromosomal translocations.

26

Immunoglobin gene rearrangements

a specialized process that utilize NHEJ to rejoin double strand breaks created by the RAG complex at specific locations in the genomes of lymphoid cells. This process results in antibody diversity in the immune system.

27

Proteins that prevent unwanted recombination events

-PARP/sing strand breaks
-Ku
-DNA dependent protein kinase
-BRCA
-ATM

28

Bloom's syndrome

Chromosome breaks

29

Ataxia-telangiectasis

Lymphoma

30

Fanconi's anemia

Leukemia