Molecular Cell biology-Drawing structures Flashcards Preview

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Flashcards in Molecular Cell biology-Drawing structures Deck (18)
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1
Q

Draw the cell undergoing prophase.

A

Refer to notes, must have centrosomes, mitotic spindle (+ends showing optional), condensing chromosomes, intact nuclear envelope, kinetochores showing. (motor proteins optional) (if the motor proteins are shown, dynein should be associated with cell membrane and pulls on astral MTs, kinesin between interpolar MTs)

2
Q

Draw the cell in prometaphase.

A

Must show NEB, (fragments of nuclear envelope), centrosomes should be at both poles, mitotic spindle showing, interpolar, astral and some kinetochore MTs. (Show dynein and kinesin 14)

3
Q

Draw the cell in metaphase.

A

All chromosomes have kinetochore-MT attachments, fragments of nuclear envelope, centrosomes at opposite poles.

4
Q

Draw the cell in anaphase.

A

Anaphase A. Show shortening of MTs attached to kinetochores and chromosomes moving to opposite poles.
Anaphase B. Show the effects of dynein and kinesin 5. (kinesin 5 force on the interpolar microtubules, interpolar MTs also elongate. Dynein pulls centrosomes to opposite poles.)

5
Q

Draw or just write (as a flow chart) the experiment for the rescue by complementation of S. pombe cdc2-ts. Starting from whole DNA sequence.

A

Show the steps of DNA library. (DNA –> RNA–> mRNA –> RT –> cDNA –> clone into plasmids –> cDNA library.
From library –> transform plasmids into plated ts-cdc2 mutants grown at restrictive temperature –> plate at restrictive –> one colony survives –> isolate plasmid –> determine sequence and protein.

6
Q

Show the in-vitro kinase (histone H1) assay that they used to determine the cdc28 kinase activity.

A
IP using anti-cdc28
add radioactive gamma-p32-ATP
add purified histone H1
incubate
SDS-PAGE
radioactive band at 29kDA
7
Q

Draw the MPF activity against location in meiosis chart.

A

Refer to notes.

8
Q

Draw the activation of any type of cdk.

A

inactive –> binding to cyclin partner (partially active) –> phosphorylation of T-loop –> CKI

9
Q

Draw the activation process of APC-cdc20 and its effect on the M-cdk. (Do not show mad2 and SAC)

A

refer to notes.

10
Q

Draw the regulation of mitotic onset.

A

refer to notes, (cdk from inactive to active with positive feedback loops)

11
Q

Draw the mitotic spindle, including motor proteins and all types of MTs.

A

refer to notes

12
Q

Draw the general process of kinetochore capture.

A

Refer to notes

13
Q

Draw the different kinetochore-MT attachements. Also show the activity of aurora B.

A

refer to notes

14
Q

Draw the pathway for the destruction of the cohesin complex.

A

Refer to notes.

15
Q

FISH experiment.

  1. How many dots for wt?
  2. How many dots for APC mutant?
  3. How many dots for securin mutant?
  4. How many dots for APC/securin double mutant.
  5. Separase mutant.
  6. For all of these, if scc1 was labeled, where would it show up?
A

Refer to notes

16
Q

Draw the SAC. (only one chromosome needs to be shown, include both APC and cdk1)

A

Kiss and run model.

17
Q

Draw the RhoA activation leading to assembly of contractile ring.

A

refer to notes.

18
Q

Draw a diagram showing the localization of actin, MHC (myosin heavy chain), RhoA, ECT2, CYK4

A

refer to notes.