Mitosis Flashcards
What are the phases of the cell cycle?
-
Interphase
- G1 phase
- S phase (DNA replication)
- G2 phase
-
Mitosis
- Mitosis (nuclear division)
- Cytokinesis (cytoplasmic division)
What controls the cell cycle?
The cell cycle is critically controlled by cyclin dependent kinases.
Describe CDK.
- CdK has 2 units.
- Cyclin + cyclin-dependent kinase.
- Cyclin does not actively participate in the cataylic reaction but it helps the enzyme.
Describe the activity of CDK during the cell cycle.
- Activity is cyclical
Describe the regulation by phosphorylation / dephosphorylation of CDK.
- Cyclin binds to CDK and it becomes an inactive cyclin-CDK complex.
- Protein kinases cause the CDK to bind an inhibitory phosphate and an activating phosphate.
- In this form it is still an inactive cyclin-CDK complex.
- An activating protein phosphatase changes this to an active cyclin-CDK complex.
Where do the 2 distinct cyclin:CDK complex control events occur in cell division?
- Active S-Cdk between G1 phase and S phase.
- Active M-Cdk between G2 phase and M phase.
- Mitotic cyclins accumulate late in the cell cycle and are active for a short period during mitotic division.
- Other cyclin:Cdk activity much earlier in the cell cycle - S-Cdk.
What is the main cyclin in the early cell cycle (G1 phase)?
What does it interact with?
- Cyclin D
- Interacts with either Cdk4 or Cdk6
What is the main cyclin during late G1 and early S phase?
What does it interact with?
- Cyclin E
- Interacts with Cdk2
Cdk2 can bind to 2 different cyclins during G1 - S phase.
What are these 2 cyclins?
- Cyclin E
- Cyclin A
What is the main cyclin during S-G2 phase?
What does it interact with?
- Cyclin A
- Interacts with either Cdk1 or Cdk2.
What is the main cyclinduring mitotic division?
What does it interact with?
- Cyclin B
- Interacts with Cdk1
How do different cyclins become destroyed?
- Cyclins become destroyed by the addition of ubiquitin, which is a small protein (76 amino acid) onto the cyclin subunit.
- It DOES NOT attach to the cyclin-dependent kinase.
- When cyclin is targeted by ubiquitin it is targeted to the proteasome and destroyed.
- The enzyme cannot work without the cyclin co-factor.
- The destruction of the cyclin conributes to the inactivation of the cyclin-dependent kinase.
Describe ubiquitination.
- 3-step process
- Initially, ubiquitin is attached to a ubiquitin-activating enzyme (E1).
- There are just 2 E1 enzymes.
- The attachment of ubiquitin is consuming energy, so there is a high energy bond between ubiquitin and E1.
- The ubiquitin is then transferred onto E2.
- There are ~35 E2 enzymes. They are known as ubiquitin-conjugating enzymes.
- Ubiquitin has many different substrates, so the Cdk:cyclin breakdown reaction has to be specific.
- The specificity comes from the E3 enzymes. These are known as ubiquitin ligases.
- These facilitate the transfer of the activated ubiquitin onto the substrate.
- Often, ubiquitination is through a lysine residue of the substrate (but not exclusively).
- Once the cyclin is ubiquitinated it can be recognised by the proteasome.
Describe the form of a proteasome.
- When a protein has been ubiquitinated, it can be recognised by a proteasome.
- Proteasome is a big complex.
- Core particle with a regulatory particle on top (regulatory particle is like a lid).
- There is a hole in the bottom.
- When the conditions are right and the ubiquitin is recognised, the ‘lid’ is opened (regulatory particle) and the protein for destruction moves inside the proteasome.
- Inside, the protein is chopped up into its composite amino acids, and discharged out through the hole in the bottom.
Describe the 2 checkpoints in G1 phase.
- Damaged DNA - the cell cannot proceed to replication if there is damaged DNA. This must be repaired before the cell can proceed to repication.
- Unfavourable extracellular environment - the cell cannot proceed if the conditions are not right.