Cell cycle Flashcards

1
Q

Which CDK binds with Cyclin D and in which phase is it expressed?

A

CDK4 and CDK6
G1-phase

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2
Q

Which CDK binds with Cyclin E and in which phase is it expressed?

A

CDK2
G1-phase

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3
Q

Which CDK binds with Cyclin A and in which phase is it expressed??

A

CDK2
S-phase

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4
Q

Which CDK binds with Cyclin B and in which phase is it expressed?

A

CDK1 aka CDC2
G2 and M-phase

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5
Q

Why do cells divide? (what are the cons for being bigger but less rather than smaller but more)

A

> Lower ability to absorb nutrients and remove waste products
Less gas exchange
Cells less likely to respond to extracellular signalling

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6
Q

Describe flow cytometry? (5 marks)

A
  1. Labelled cells suspended in fluid and passed through single file
  2. laser light focussed on cells
  3. Excites any flourescence detected
  4. emitted flourescence detected
  5. Scattered light detected—> measures total cell number.
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7
Q

What is the amount of flouresence directly proportional to in flow cytometry?

A

Amount of DNA
More DNA = more intercalation = more flourescence

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8
Q

What is Flow cytometry used for? (there are 4 uses)

A

Cell counting/sorting
Protein expression
Apoptosis
Cell cycle analysis

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9
Q

What are the 3 common methods to study the cell cycle?

A
  1. Light/time lapse microscopy—> cellsin mitosis are rounded up
  2. DNA binding dyes BrdU, incorporate into new DNA formed in S-phase
  3. Flow cytometry
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10
Q

What is the function of G1?

A

Increase the size of the cell, duplicate organelles, PREPARE FOR DNA SYNTHESIS

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11
Q

What is the function of E2F?

A

TF, stimulates expression of genes needed for the cell cycle

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12
Q

What is Rb?

A

Transcriptional repressor, blocks the function of E2F

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13
Q

What are the 2 ways Rb inhibits E2F and describe each?

A

Passive Repression—> Rb binds E2F and prevents it activating transcription of genes nedded for S-phase
Active repression—> Rb recruits the Sin3 & HDAC proteins, Blocks transcription by modifying chromatin

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14
Q

What is the function of HDAC?

A

HDAC= histone DeAcytylases
Deacetylates histones- closes up chromatin and blocks transcription
Stops polymearse to bind to DNA so no transcription occurs.

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15
Q

Describe the Removal of Active Repression. (4)

A
  1. Cyclin-D-CDK4 or 6 —>phosphorylates Rb
  2. Blocks Sin3 binding to Rb
  3. HDAC—> repression removed=transcription
  4. expression of subset of genes needed for late G1 and early S-phase
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16
Q

Describe the Removal of Passive Repression (3)

A
  1. Cyclin E-CDK2—> 2nd Rb phosphorylation
  2. Blocks Rb binding E2F—> removes passive repression of E2F
  3. Expression of genes needed to complete S-phase:histones, cyclin B, nucleotide synthesis for new DNA
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17
Q

What is the G1 restriction Checkpoint?

A

Point of no return, checks DNA integrity

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18
Q

What has to happen for the cell to go through G1 checkpoint?

A

Cells must inactivate: Rb

Rb —> Transcriptional repressor blocks the function of E2F

E2F —> TF, stimulates expression of genes needed for the cell cycle

19
Q

What cyclins and CDK are in the G1 phase?

A

Cyclin D + CDK4/6
Cyclin E + CDK2

20
Q

What cyclins and CDK are in the S?

A

Cyclin A + CDK2

21
Q

What cyclins and CDK are in the G2 + M?

A

Cyclin B + CDK1

22
Q

What do CKIs do?

A

Inhibit cell cycle progression

23
Q

What is active repression by Sin3-HDAC

A

DNA —> tightly packed around histones

Transcription—> DNA opened up allows polymerase access

Requires histone acetylation

HDACs—> Histone DeAcetylases

De-acetylates histones= closes up chromatin and blocks transcription

24
Q

What is Ori?

A

Region of DNA where replication is initiated
Eukaryotes: have multiple Ori’s
Prokaryotes only 1 Ori cause it has a circular chromosome

25
How is DNA replication controlled? THERE ARE POINTS FOR EACH STAGE (G1/S)
G1: ORC vinds to Ori After initiation of cell cycle: E2F partially activated->active repression by Rb Allows expression of E2F target gene CDC6 which binds to ORC CDC6, ORC & CDT1 recruit inactive 2x MCM helicase Binding of MCM helicase to DNA forms the pre-replication complex At G1/S transition: Cyclin C-CDK2 phosph thepre-replication complex DNA Helicasesunwind DNA--->inactive DNA polymerase recruited S: Cyclin A-CDK phosphorylates--> DNA polymerase, CDC6, ORC Polymearse activated, ORC inactivated->dissociates from DNA, CDC6 targeted for destruction->breaks link between Ori and DNA polymerase
26
What drug is used to improve elderly health?
Senolytic drugs: kill senescent cells, as SASP is associated with age related disease such as type 2 diabetes, alzheimers etc.
27
What is a kinetochore?
Large protein structure built at the centromere allows attachment of the spindle microtubules
28
What is the role of Cyclin B?
B controls G2 and mitosis: CYTOPLASMIC RETENTION SEQUENCE CRS NUCLEAR LOCALISATION SEQUENCE NLA CRS is dominant over the NLS G2 progresses= increased cyclin B
29
How is Cyclin B-CDK1 activated?
Inactive: phosphorylated by Wee1, Cyclin B-CDK1 is bound by CRS CRS phosphorylation by PLK-1 and ERK blocks cytoplasmic retention Cyclin B-CDK1 enters nucleus remains inactive PLK-1 phosphorylates and activates CDC25 CDC25 removes CDK1 inhibition Active cyclin B-CDK1 phosphorylation and furtheractivates CDC25 Positive feedback on cyclin-B-CDK1 Cyclin B-CDK1 can now REGULATE PROGRESSION THROUGH MITOSIS
30
What is Cohesin?
Forms a ring like structure around the sister chromatid
31
What is condensin?
Restructures and condenses the DNA
32
What does Cyclin B-CDK1 do in late G2 and prophase?
Condensin need to be activated SL1 needs to be inactivated Cyclin B-CDK1 phosphorylates both. condensin activates, SL1 deactivates Promotes chromosomal condensation
33
What does Cyclin B-CDK1 phosphorylate in prometaphase?
Lamin- causes nuclear membrane to breakdown
34
What is Upiquitin and what are the 3 classes?
Protein degradation 3 classes: E1 activates ubiquitin E2 receives Ubq from E1 and prepares it for ligations E3 ligated Ubq to target proteins
35
Cyclin B-CDK1 function in metaphase?
Phosphorylates APC and Cdh1 swapped for CDC20 Phosphorylates Seperase and securin binding
36
What happens in anaphase?
APC-CDC20 ubiquinates securin releasing seperase Seperase ubiquitinates cohesin (sister chromatids seperate) Seperase ubiquitinates Fear and MEN- activates CDC14b APC-CDC20 ubiquitinates Cyclin-B
37
What happens in Telophase/cytokinesis?
CDC14b is the main factor!! Dephosphorylates Cdh1 and APC --> swap of CDC20 for Cdh1, so APC-Cdh1 and CDC20 alone?!---> Cyclins Ubiquitinated and destroyed CDC14b Dephosphorylates myosin--> contractile ring seperates 2 daughter cells CDC14b dephosphorylated Rb->represses E2F CDC14b dephosphorylates SL1 so transcription restarts CDC14b dephosphorylates lamin--> nuclear mem reassembly---> CDC14b --> sequestered in newly forming nucleolus
38
What happens in the resriction checkpoint(G1)? HOW IS THE CELL CYCLE ARRESTED WHAT HAPPENS WHEN THE DNA IS REPAIRED
ATM detects DNA damage causes ATM to become phosphorylated and seperate (they are in pairs). ATM phosphorylates ChK2-->INDUCE AND STABALISE p53 and inhibit MdM2 is a ubiquitin ligase which targets p53 p53 levels increase---> p21 induced which is a cyclin dependant kinase inhibitor--->Cyclin E-CDK2 inhibited preventing full activation of E2F and preRC CAUSING THE CELL CYCLE TO ARREST DNA repaired ChK2 is inactivated so MdM2 is no longer repressed so p53 is ubiquinated--> decrease of p53 and p21 so cyclin E-CDK2 in repressed so entry to S-phase
39
What happens in the G2/M checkpoint?
DNA damage activates ATR which phosphorylates ChK1 which inhibit CDC25 and activates Wee1 Wee 1 inhibits Cyclin B-CDK1 CDC25 activates Cyclin B-CDK1 CELL CYCLE ARRESTS IF mutation on wee1 then cell cycle is unregulated: cancer etc.
40
What happens in the metaphase checkpoint?
Negatively regulates APC-CDC20---> Cyclin B-CDK1 phos Cdh1 and APC APC dissociates from Cdh1 and binds CDC20 APC-CDC20 ubiquitinates securin MCC bind and inhibit APC-CDC20 MAD proteins sense if spindle fibre is not correctly positioned etc. When the sister chromatids are correctly loaded onto the mitotic spindle MAD and BUB will dissociate from APC-CDC20 so they can ubiquinate Securin
41
What is the intrinsic apoptosis pathway?
Bcl regulate VDAC and ANT channels anti apoptotic: Prevent pore opening and cytochrome c leaving the mitochondria Pro apoptotic: keep pores open and allow cytochrome c to leave the mitochondria BAX allows cytochrome c to leave throught the VDAC (opening it) Bcl2 stops VDAC to open for cytochrome c In high levels BAX can let cytochrome c to leave on it own without VDAC
42
What happens once cytochrome c is in the cytoplasm?
Cytochrome c and APAF bind together This causes a cleavage in Caspase 9 to activate it This activates a cascade to Caspase 3/or 6 caspase 3 can induce apoptosis or caspase 7 and then apoptosis Caspase 6 only induces apoptosis
43