Lecture 9 Genome Instability and Checkpoints (WIP) Flashcards Preview

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Flashcards in Lecture 9 Genome Instability and Checkpoints (WIP) Deck (32)
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1

What is meant by genome instability

An elevated rate of genetic alterations

2

Tumorigenesis is a multi-stage process T or F

T

3

Give some examples of checkpoints throughout the cell cycle

Restriction point that governs entry into S phase from G1 intra-S-phase checkpoint that checks for DNA damage or stalked replication forks G2M phase checkpoint that checks that all DNA has been replicated and then the metaphase to anaphase transition in mitosis that checks whether all chromosomes are attached to the spindle

4

Why is the intestinal epithelium used as a model for the changes in cancer cells over time

This is the best characterised and understood cancer in terms of the chromosomal abnormalities that correspond with its progression

5

Outline the progression of intestinal epithelium towards tumorigenesis

Normal epithelium hyperplastic epithelium early/intermediate/late adenoma carcinoma metastasis

6

Below is a diagram outlining the progression of intestinal epithelium during tumorigenesis. Fill in above the arrows with the specific chromosomal abnormality that causes each transition

Loss of APC DNA hypomethylation activation of KRas loss of Smad4 loss of p53

7

Give examples of some of the chromosomal changes that occur in the progression of cells during tumorigenesis

Loss of tumour suppressor genes proto-oncogene to oncogene transitions and changes in epigenetic regulation of the genome

8

What is meant by a driver mutation

A mutation that emerges early in cancer and occurs in a large majority of all cancers of a certain type

9

There are over 15 different driver mutations in colorectal cancer give some examples of these

APC PIK3CA KRas TP53 FBXW7 β-catenin axin-2

10

Why are driver mutations better therapeutic targets in the development of cancer treatments

Because mutations in these genes are extremely common and occur early so this will maximise the patients who could benefit and would mean that the cancer is stopped early in its progression

11

What is significant about the various different mutations that occur at each point in tumorigenesis

Mutations at each stage in tumorigenesis are often components of the same pathway

12

What hypothesis was proposed to explain how tumour cells manage to accumulate mutations

The mutator hypothesis was proposed to explain the accumulation of all the mutations in tumour cells. This is the idea that tumour cells acquire initial mutator gene mutations that then increases the rate of subsequent mutations

13

What are the two type of instability seen in cancer cells and how do they differ

Chromosomal instability – chromosomal rearrangements and aberrations as well as loss of heterozygosity aneuploidy/polyploidy and gene amplifications. Microsatellite instability – point mutations and base substitutions as well as microdeletions or insertions giving rise to missense mutations

14

Microsatellite and chromosomal instabilities are common in normal cells T or F

F – they are extremely rare occurring once in every 10million cell divisions

15

Where is it that mutator genes are often found

Form part of the DNA repair systems

16

What are the different methods of DNA repair

Base excision repair nucleotide excision repair homologous recombination and non-homologous end joining

17

What mechanisms exist to prevent DNA replication and processing errors

Precursor control polymerase proof-reading and mismatch repair

18

What is meant by integral dependency. How does this relate to the cell cycle

Integral dependency is where one component is reliant on the previous component. It suggests that in order for the cell cycle to progress each phase needs to be completed before the next one can begin

19

Give an example of integral dependency

Bacteriophage assembly – whereby one component of virus assembly depends on the previous component. In other words the way in which the virus assembles is inherent to the structure of the virus

20

What is the effect of RAD9 mutations in the growth of S. cerevisiae

Cells lacking RAD9 are healthy in the absence of extrinsic interference. Hence the growth curves of RAD9 and rad9 yeast cells are the same

21

What were the results of experiments carried out with yeast harbouring temperature-sensitive cdc9 mutations

FACS analysis of cdc9 mutants at permissive temperature looks normal however FACS analysis of cdc9 mutants at a restrictive temperature indicates G2 arrest

22

What is the role of cdc9 and how does this account for what is seen in the temperature-sensitive mutants

Cdc9 codes for a DNA ligase that is important in lagging strand synthesis where it is required to join Okazaki fragments during DNA replication. If you lack cdc9 there will be complete DNA synthesis but this DNA will be fragmented. Hence cdc9 mutants at restrictive temperatures arrest after S phase as all of the DNA is synthesised but is fragmented so that M phase cannot proceed

23

What is meant by cell viability how does it differ from cell cycle arrest

Viability is the ability of cells to grow and make daughter cells. It isn’t related to cell cycle arrest as in the case of temperature-sensitive mutants changing the permissive temperature of a cdc mutant back to a permissive one will result in the cells coming out of arrest and therefore being viable

24

Describe what these results obtained by Lee Hartwell show about the viability of cdc9/rad9 mutants

These graphs show that wild type yeast as well as cdc9 and rad9 mutants are all viable a hence have a near 100% viability. However the rad9/cdc9 combination mutants have drastically reduced viability. Interestingly the longer you leave the rad9/cdc9 cells the more rapidly that viability is reduced. This implies that cells undergoing replication stress such as due to non-functional DNA ligase/cdc9 require RAD9 in order to maintain their viability. Once this is also lost cells are no longer viable

25

How can loss of viability be used as a screen for checkpoint mutants

You can identify which mutations in cell cycle genes cause a loss of viability when crossed with rad9 mutants

26

What kind of gene was RAD9 identified to be

A checkpoint gene

27

What is meant by a checkpoint gene

An extrinsic control gene that ensures that viability can be maintained when an extrinsic stress is applied

28

What is meant by mitotic catastrophe

Where cells attempt mitosis when they shouldn’t

29

What is premature chromosome condensation

A phenotype associated with a loss of the S/M phase checkpoint. This is indicative of mitotic catastrophe where cells have attempted to enter mitosis without completion of S phase

30

How has premature chromosome condensation lead to the identification of a checkpoint gene involved in the S phase to mitosis transition

BHK cells were mutagenized so as to harbour temperature-sensitive cell cycle mutations. These cells were the placed at the restrictive temperature before being treated with a drug used to stop cells in mitosis so as to look at the chromosome condensation. This lead to the identification of premature chromosome condensation indicative of a checkpoint gene