Lecture 4 Flashcards

1
Q

At which two points can the cell cycle be stopped if genome is damaged?

A

Restriciton point: Blocks passage into S phase

Intra-S phase: Blocks DNA replication

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
2
Q

What three routes can a cell take after checks for DNA damage at restriciton point?

A

Divide - enter S phase

Arrest - pause to rest

Die - undergo apoptosis

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
3
Q

Which gene product is commonly compromised in cancer cells and hence cause the cell to loss ability to deal with DNA damage ‘decisions’?

A

P53

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
4
Q

Which type of mutations ofP53 are commonly seen?

A

Alterations in arganine248, which binds the DNA backbone, and other DNA binding residues

Mutations in the zinc finger motif, which is vital for the interaction between loop 2 and 3 of the p53 to allow DNA binding.

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
5
Q

Which Cdki is regulated by P53?

A

p21 is upregulated by P53

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
6
Q

What role does p21 play?

A

Inhibits Cdk2 (entry into S phase)

Inhibits PCNA (halts ongoing DNA replication)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
7
Q

What are the roles of PCNA?

A

DNA polymerase ‘sliding clamp’

Holds polymerase onto the DNA and increases possible length of product by several orders of magnitude

Also attracts DNA repair factors to arrested DNA replication forks (not inhibited by P21)

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
8
Q

What is the other way a cell can deal with damage if it has lost its p53?

A

The DNA damage response in S1

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
9
Q

Why is the DNA damage response being targeted as a possible cancer therapy?

A

If cancer cell has lost p53 dependent cell cycle arrest then knocking out DNA damage response should cause cell death. But healthy cells will be unaffected due to still having p53 pathway available

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
10
Q

What are the four broad classes of molecule involved in DNA damage response?

A
  1. Sensors
  2. Mediators
  3. Tranducers/Effectors
  4. Responders
How well did you know this?
1
Not at all
2
3
4
5
Perfectly
11
Q

What is the main role of the transducer molecules?

A

Amplify the once local signal to spread across the entire genome

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
12
Q

Outline the general cascade for recruiting repair proteins to sites of damage

A

A PI3 Kinase (PIK) family kinase is recruited to the site of damage by partner proteins where they phosphorylate signal proteins to recruit repair proteins

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
13
Q

Outline an example cascade for recruiting repair proteins to sites of DS break damage

A

Nbs1, part of the MRN complex, recruits ATM which phosporylates CHK2 and causes signalling.

CHK2 can then phosphorylate cdc25A leading to its destruction and inhibiting its ability to remove the inhibitory phosphate group from cdk2, causing cell cycle arrest.

Additionally AMT and the MRN complex can then cuase DNA repair through a variety of mechanisms

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
14
Q

How will ATM ‘choose’ between homologous repair or non-homologous end joining?

A

NHEJ is the default pathway but HR will be used if in S phase - measures the level of CDK which are higher in S phase

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
15
Q

Why does CHK2 offer a quick damage response?

A

Constantly available in the genome and does not need to be made when damage discovered

How well did you know this?
1
Not at all
2
3
4
5
Perfectly
16
Q

What is activated by replicative stress?

A

CHK1 via ATR

17
Q

Germ line muations of what protein are found in familial breast and ovarian cancers?

A

BRCA1

50% for breast

70% for ovarian

18
Q

What is the role of BRCA1?

A

Scaffolding protein for all proteins involved in damage repair

19
Q

How is damaged flagged by ATM/ATR?

A

Phosphorylation of which causes ubiquitination

20
Q

What non-repair protein is attracted by this H2AX flag?

A

Cohesin - which acts ot hold the chromatids togther

21
Q

What is the outcome of chronic activation of DDR?

A

Deactivation and then hyper mutation

22
Q

What can cause replication forks to slow down or stall?

A
  • Disruption of nucleotide pools
  • DNA damage/obstructions
  • DNA secondary structures stop unwinding of DNA
23
Q

Why would a stalled replication fork be dangerous?

A

Transiently in single stranded form