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Microbiology 2 > Test 3: Cell Cycle > Flashcards

Test 3: Cell Cycle Flashcards

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Biology 101 flashcards
1
Q

4 parts of Cell Cycle with identifiers & n

A
  1. G1: (1n): cell grows
  2. S: (>1n): DNA rep
  3. G2: (2) Prepares for mitosis
  4. M: (2n) Mitosis
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2
Q

Measuring Cell Cycle: 1. Determine Phase of the cell cycle is in. Check for S-phase by…

A

DNA replication only in S phase, so Thymedine-label (T in DNA) fed to cell for short period, washed, and look for cells with label

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3
Q

Measuring Cell Cycle: 1. Determine Phase of the cell cycle is in. Check for M-phase by…

A

Condensed chromosome

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4
Q

Measuring Cell Cycle: 1. Determine Phase of the cell cycle is in. Test for G1 by…

A

Cytometer, given it is only phase with 1 n cell. Label DNA with fluorescent probe.

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5
Q

Measuring Cell Cycle: 2. Measure length of each phase by…

A

% of cells in phase = % that phase takes up in cell cycle

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6
Q

Frog Oocyte Experiment: F. oocytes chosen because (4)

A
  1. large cell, easy to inject
  2. lots of cytoplasm, lots of pfoteins to be purified
  3. lots produced
  4. FROZEN IN G2 PHASE UNTIL SPERM ENTERS
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7
Q

Frog Oocyte Experiment: G2 phase frozen because either … (2)

A
  1. missing something needed to go thru cell cycle

2. something is inhibiting cell cycle

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8
Q

Frog Oocyte Experiment: To determine effect of G2 phase arrest, took oocyte and injected cytoplasm of cycling cells. Which phase of injected cell was effective? What was the conclusion?

A

M - oocyte starts mitosis. Hence, something is in M phase that triggers G2 -> M called Mitosis Promoting Factor (MPF)

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9
Q

Frog Oocyte Experiment: MPF exists in .. phase. It is highly…

A

M-phase only; conserved

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10
Q

Fission Yeast Experiment: Chosen because (4)

A
  1. Single Celled
  2. Easily Grown
  3. Haploid, hence easy genetics
  4. Only 2 cell cycle phenotypes
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11
Q

Fission Yeast Experiment: 2 cell cycle phenotypes were…

A
  1. arrested phenotype: won’t go through cell cycle and needs activator i.e. MPF
  2. wee phenotype: cycles too quickly, removed an inhibitor of sort, hence very small/no growth
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12
Q

Fission Yeast Experiment: Temperature-sensitive mutant has 2 phenotypes according to temperature. phenotype name and temperature corresponding to it?

A
  1. WT Phenotype /can cycle at permissive T

2. mut phenotype/wee or arrested at restrictive T

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13
Q

Fission Yeast Experiment: cdc2 is…. that unlike expected…

A

T-sensitive mut protein kinase that arrested at G2 like oocyte BUT did not react with MPF

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14
Q

Fission Yeast Experiment: cdc2 must be activated by…

A

cyclin

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15
Q

Fission Yeast Experiment: cyclin regulates kinase activity of cdc2 by forming… cdc2 is hence called…

A

cyclin:cdc2 complex; cyclin-dependent kinase

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16
Q

Fission Yeast Experiment: when cyclin:cdc2 complex is injected, oocyte…. Conclusion?

A

triggered m-phase. hence, the complex is GFP

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17
Q

Fission Yeast Experiment: level of cyclin v cdc2 across various cell cycles

A

cdc2 constant; cyclin peaks slowly to M and drops as soon as it ends, to rise again slowly from G1 to S to G2

18
Q

cdc regulation: Upon further scrutiny, cdc2 + cyclin by itself does not wholly explain MPF activity. cak, wee1, and cdc25 is involved. explain.

A

G2: cak- & wee1-cdc2:cyclin:
cak phosphorylates cdc2 on Thr residue (act)
wee1 phosphoorylates cdc2 on Tyr residue (inhibit)
M: cdc25 removes phosphate from Tyr (act)

19
Q

T/F cdc regulation: cdc25 is necessary for activation of GPF

A

T

20
Q

cdc regulation: MPF has feedback loop that activates… which ….

A

activates cdc25, which deactivates effect of wee1 by dephosphorylating Tyr

21
Q

cdc regulation: MPF’s sharp peaks of activity explained by…

A

self-propagation

22
Q

cdc regulation: cak- mutation leads to

A

no cycling; G2

23
Q

cdc regulation: wee1- mutation leads to

A

++ cycling; wee

24
Q

cdc regulation: cdc25- mutation leads to

A

no cycling; G2

25
Q

cdc regulation: wee1- and cdc25- mutation leads to…

A

++ cycling; wee

26
Q

MPF: is a kinase. it effects 4 things

A
  1. chromosome condensation
  2. nuclear envelope breakdown
  3. mitotic spindle formation
  4. cyclin degradation
27
Q

MPF: 1. chromosome condensation… by

A

phosphorylating histones and condensins

28
Q

MPF: 2. nuclear envelope breakdown… by

A

phosphorylating lamin (which nuclear envelope is made of to prevent protein protein interaction

29
Q

MPF: 3. mitotic spindle formation… by

A

dynamic instability changed by +++ ++ MAPs associated with microtubules; phosphorylates MAPS

30
Q

MPF: 4. cyclin degradation… by

A

end of mitosis to decrease cyclin steeply

31
Q

Cyclin:CDK is common mode of regulation for cell cycle. … ocntrols entry into S-phase from G1

A

cdc2:G1 cyclin

32
Q

Cyclin:CDK: G1 -> S entry point critical b/c

A

Chromosome duplicates in S-phase, commits to mitosis after that

33
Q

G1 Cyclin:Cdc2: cyclin substrate is…

A

Rb, which inhibits transcription by binding to ts E2F

34
Q

G1 Cyclin:Cdc2: 1 cyclin, Rb, E2F. Relate.

A

Rb:E2F prevents transcription.
G1:cc2 phosphorylates Rb
E2F now free for transcription of genes for S-phase

35
Q

Cyclin:CDK: S-phase itself trigged by…

A

S-Cyclin:Cdc6

36
Q

S-Cyclin:Cdc6: S-cyclin… (2)

A
  1. initiates DNA replication

2. Prevents re-replication of DNA

37
Q

S-Cyclin:Cdc6: Relate cdc6, Origin Recognition Complex (ORC), ori, and S-Cdk

A
  1. ORC is bound to ori
  2. cdc6 binds to ORC to recreate pre-replicative complex
  3. S-cdk triggers phosphorylates cdc-6 & triggers S-phase
38
Q

S-Cyclin:Cdc6: S-cyclin initiates DNA replication by…

A

@ ori, pre-replication complex sits on DNA and waits for S-phase. s-Cdk triggers replication as cdc6 is phosphorylated and comes off of ORC

39
Q

S-Cyclin:Cdc6: S-cyclin prevents rereplication by…

A

s-Cdk phosphorylates cdc6, which is soon degraded. cdc6-P triggers its own destruction so it does not form more pre-replicative complex

40
Q

DNA Damage: when damaged, cell…

A

cell arrests to give time for repair

repair

41
Q

DNA Damage: relate p53 and p21

A

DNA damage leads to + transcription of p53 for DNA repair

leads to + transcription of p21 for cdk inhibition

Microbiology 2 (8 decks)

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