Garantie de satisfaction à 100% Disponible immédiatement après paiement En ligne et en PDF Tu n'es attaché à rien
logo-home
Recherché précédemment par vous
Recherché précédemment par vous
logo
Lecture Notes on the Cell Cycle (MCB2023S) $7.54
Ajouter au panier
Accédez rapidement à
Aperçu
  2.  
  3. University Of Cape Town (UCT)
  4.  
  5. MCB2023S

Notes de cours

Lecture Notes on the Cell Cycle (MCB2023S)
 0 fois vendu
  • Cours
  • MCB2023S
  • Établissement
  • University Of Cape Town (UCT)

Comprehensive Lecture Notes on the Cell Cycle, however some of the experimental procedures are all documented well but the findings from the experiments are all present. Contents can be seen on the first page. Summary is that it includes: - Regulation of the G2/M checkpoint - Regulation...

[Montrer plus]

Aperçu 4 sur 34  pages

Infos sur le Document

  • 11 novembre 2021
  • 34
  • 2021/2022
  • Notes de cours
  • Nicola illing
  • Toutes les classes

Sujets

  • p53
  • cancer
  • mitosis
  • meiosis
  • cyclin
  • mpf
  • cell cycle
  • sea urchin
  • cdk
  • checkpoint
  • regulation
  • wee1
  • cdc25
  • dosage effect
  • cell division
  • cell division cycle
  • phosphorylation
  • budding yeast
  • fission yeast
  • s pombe
  • s

École, étude et sujet

  • University of Cape Town (UCT)
  • MCB2023S
Tous les documents sur ce sujet (11)

Vendeur

avatar-seller
mattclothier13

Avis reçus

Aperçu du contenu

Cell Cycle
CELL CYCLE.................................................................................................................................................... 1
INTRODUCTION............................................................................................................................................ 3
TRADITIONAL METHODS OF STUDYING THE CELL CYCLE..................................................................................................3
MODEL SYSTEMS FOR STUDYING THE CELL CYCLE..........................................................................................................3
PHASES OF THE CELL CYCLE......................................................................................................................................3
DIFFERENT TYPES OF CELL CYCLES.............................................................................................................................3
CHECKPOINTS IN THE CELL CYCLE..............................................................................................................................4
REGULATORS IN CELL CYCLE....................................................................................................................................4
Dominant M-phase Factor:............................................................................................................................4
Dominant S-phase factor:...............................................................................................................................4
Block to DNA re-replication:...........................................................................................................................5
REGULATION OF THE G2/M CHECKPOINT...................................................................................................... 5
IDENTIFICATION OF MPF........................................................................................................................................5
Discovery of Maturation Promoting Factor (MPF): the dominant M-phase factor.......................................5
Polar Bodies....................................................................................................................................................6
IDENTIFICATION OF MITOTIC CYCLIN..........................................................................................................................6
GENETIC ANALYSIS OF THE CELL CYCLE......................................................................................................................7
Isolation of cell division cycle (cdc) mutants..................................................................................................7
Characterization of cell cycle mutants...........................................................................................................7
Cloning by complementation of cdc2ts, wee1ts, cdc25ts and cdc13ts mutants.................................................7
What did the cloning tell us?.....................................................................................................................................8
Cloning of wee1, cdc25 & cdc13.....................................................................................................................8
MITOTIC PROMOTION FACTOR (MPF) = P34CDC2 & CYCLIN B.........................................................................................9
CELL CYCLE.................................................................................................................................................... 9
Raising antibodies to cdc2..............................................................................................................................9
i. Does the anti-cdc2 antibody cross-react with a protein in the partially purified Xenopus MPF fraction?..9
ii. Do any other proteins in the MPF fraction co- immunoprecipitate with the anti-cdc2 antibody?............9
Mechanism of co-immunoprecipitation assay:.........................................................................................................9
Results:.........................................................................................................................................................10
iii. Does the co-immunoprecipitate have MPF activity?...............................................................................10
Results:.................................................................................................................................................................... 10
DOSAGE EFFECT OF WEE1, CDC25 AND CDC2 ON THE CELL CYCLE................................................................................10
Results:.........................................................................................................................................................11
ROLE OF CYCLINB................................................................................................................................................ 11
In Vitro assay for cell cycle...........................................................................................................................11
Findings:.................................................................................................................................................................. 11
Is the mitotic cyclin B required for MPF activity?.........................................................................................12
Is cyclin B degradation required for exit from the cell cycle?.......................................................................12
Regulation of cyclin B degradation..............................................................................................................12
PHOSPHORYLATION REGULATION:...........................................................................................................................13
Experimental Questions:...............................................................................................................................13
Experimental Tools:......................................................................................................................................13
Experimental procedure:..............................................................................................................................13
1. Does the phosphorylation status of Cdc2 change during the G2/M transition?......................................14
2. Which amino acids are phosphorylated on Cdc2?...................................................................................15
Assay:......................................................................................................................................................................15
Which tyrosine is phosphorylated and dephosphorylated?....................................................................................15
Inhibitory effect of Y15-P:.......................................................................................................................................15
Wee1 & Mik1 phosphorylate cdc2 (cdk1) on Y15.........................................................................................16
Analysis of mik1 and wee1 mutants........................................................................................................................16
Cdc25 dephosphorylation of cdc2(cdk1)......................................................................................................16
Assay:......................................................................................................................................................................16
WHAT ARE THE TARGETS OF ACTIVE MPF?..............................................................................................................17
SUMMARY:.........................................................................................................................................................17

, Cell Cycle
REGULATION OF THE G1/S CHECKPOINT...................................................................................................... 17
CDC2/CDC28 IS REQUIRED FOR START (THIS CHECKPOINT).........................................................................................17
Budding Yeast Life Cycle...............................................................................................................................17
IDENTIFICATION OF G1 CYCLINS:.............................................................................................................................18
Genetic/Mutant screen in S. cerevisiae........................................................................................................18
Dosage effects of G1 cyclins.........................................................................................................................18
CELL CYCLE IN S. CEREVISIAE..................................................................................................................................18
IDENTIFICATION OF MAMMALIAN CYCLINS & CDC2 HOMOLOGUES (CDK1, CDK2…CDK5)...................................................18
Cyclin D is required for the G0 to G1 transition in mammalian cells............................................................19
IDENTIFICATION OF MAMMALIAN CYCLIN-DEPENDANT KINASES (CDKS)..........................................................................19
REGULATION OF MAMMALIAN CELL CYCLE...............................................................................................................20
IDENTIFICATION & CHARACTERIZATION OF CDK INHIBITORS..........................................................................................20
Identification of p16 in association with cdk4..............................................................................................20
Assay for cdk4/cyclinD activity.....................................................................................................................20
TUMOUR SUPPRESSORS: RB & P53.............................................................................................................. 21
CDK INHIBITORS P16, P21, P27.............................................................................................................................21
CANCER GENETICS AND THE RETINOBLASTOMA GENE.................................................................................................21
Action of Rb Protein......................................................................................................................................21
P53 – MOST FAMOUS TUMOUR-SUPPRESSOR GENE...................................................................................................22
DNA DAMAGE............................................................................................................................................. 23
DNA DAMAGE RESPONSE GENES...........................................................................................................................24
DNA DAMAGE CONTROL AT G2/M CHECKPOINT.....................................................................................................24
Experimental Evidence:................................................................................................................................24
DNA DAMAGE CONTROL AT G1/S CHECKPOINT.......................................................................................................27
Question: When cells are exposed to gama-irradiation, do Cdc25A levels affect progress through the S-
phase?..........................................................................................................................................................27
P53 Independent Pathways..........................................................................................................................28
P53 Dependant Pathways............................................................................................................................28
CANCER...................................................................................................................................................... 29
........................................................................................................................................................................ 29
PROPERTIES OF CANCER........................................................................................................................................30
Multi-Hit model............................................................................................................................................30
Cancer Phenotype.........................................................................................................................................31
TUMOUR SUPPRESSOR GENES................................................................................................................................31
Properties:....................................................................................................................................................31
PROTO-ONCOGENES AND ONCOGENES....................................................................................................................31
DNA TUMOUR VIRUSES........................................................................................................................................32
CANCER TREATMENT & DIAGNOSIS............................................................................................................. 32
IONIZING RADIATION:...........................................................................................................................................32
CHEMOTHERAPY..................................................................................................................................................33
Mechanisms of action of different types of Drugs in Chemo:......................................................................33
HORMONAL TREATMENTS.....................................................................................................................................34
CANCER DIAGNOSIS.............................................................................................................................................34
COMBINE RADIATION TREATMENT WITH WEE1 INHIBITORS..........................................................................................34

, Cell Cycle
Introduction
Traditional methods of studying the cell cycle

DNA Replication
- 3H-Thymidine incorporation
o Inject autoradiated thymidine into cell then use autoradiography to see if its
been incorporated into the DNA
- Artificial Thymidine analog Bromo-deoxyuridine (BrdU)
o Readily incorporates opposite Adenines.
o If cell is replicating then BrdU will be incorporated
o Fluorescently labelled anti-BrdU antibodies will show up if cell replication has
occurred.


Model systems for studying the cell cycle
- Mammalian cells proliferating in culture
- Genetic analysis in budding yeast
- Animal embryos: Xenopus & sea urchin eggs



Phases of the cell cycle
G1 Phase:
- Immediately after mitosis
S phase:
- DNA Replication
G2 Phase:
- Before mitosis
M Phase:
- Mitosis
G0 Phase:
- After mitosis if cell leaves cycle and stops dividing
Interphase:
- G1 + S + G2
- DNA is diffusely distributed within nucleus


Different types of Cell Cycles
- Embryonic Cell cycles
o Short cycles of constant duration
o Very synchronous
- Growing cell cycles
o Longer cycles of variable duration

, Cell Cycle
- Reversible exit from cell cycle
o Some cells stop growing and leave the cycle until interacted with a growth
factor etc.


Checkpoints in the Cell Cycle




Regulators in Cell Cycle

Dominant M-phase Factor:
- 1970s: Cell fusion experiments showed that there were different regulators at
different stages of the cell cycle
- Active during Mitosis
- Synchronized mammalian cells, mixed cells in presence of virus or chemical agent
that caused their membranes to fuse- formed hybrid cells
- Results:
o G1 fused with M phase cells = chromatin immediately condensed
o G2 fused with M phase cells = chromatin immediately condensed
o S phase fused with M phase cells = chromatin immediately condensed
- Conclusion:
o Dominant M-phase factor
o Mitotic cells must contain a diffusible factors that induce mitosis when
introduced to cells at other stages of the cell cycle

Dominant S-phase factor:
- When G1 and S cells fused
- G1 cells immediately began to replicate their DNA
- Conclusion:
o Factors from S phase cell must induce DNA replication in G1 nucleus – hybrid
cell delays mitosis until replication has completed.
o S-phase factor induces DNA replication

Les avantages d'acheter des résumés chez Stuvia:

Qualité garantie par les avis des clients

Qualité garantie par les avis des clients

Les clients de Stuvia ont évalués plus de 700 000 résumés. C'est comme ça que vous savez que vous achetez les meilleurs documents.

L’achat facile et rapide

L’achat facile et rapide

Vous pouvez payer rapidement avec iDeal, carte de crédit ou Stuvia-crédit pour les résumés. Il n'y a pas d'adhésion nécessaire.

Focus sur l’essentiel

Focus sur l’essentiel

Vos camarades écrivent eux-mêmes les notes d’étude, c’est pourquoi les documents sont toujours fiables et à jour. Cela garantit que vous arrivez rapidement au coeur du matériel.

Foire aux questions

Qu'est-ce que j'obtiens en achetant ce document ?

Vous obtenez un PDF, disponible immédiatement après votre achat. Le document acheté est accessible à tout moment, n'importe où et indéfiniment via votre profil.

Garantie de remboursement : comment ça marche ?

Notre garantie de satisfaction garantit que vous trouverez toujours un document d'étude qui vous convient. Vous remplissez un formulaire et notre équipe du service client s'occupe du reste.

Auprès de qui est-ce que j'achète ce résumé ?

Stuvia est une place de marché. Alors, vous n'achetez donc pas ce document chez nous, mais auprès du vendeur mattclothier13. Stuvia facilite les paiements au vendeur.

Est-ce que j'aurai un abonnement?

Non, vous n'achetez ce résumé que pour $7.54. Vous n'êtes lié à rien après votre achat.

Peut-on faire confiance à Stuvia ?

4.6 étoiles sur Google & Trustpilot (+1000 avis)

65040 résumés ont été vendus ces 30 derniers jours

Fondée en 2010, la référence pour acheter des résumés depuis déjà 15 ans

Commencez à vendre!

Récemment vu par vous

Cas ·

(0)

BIOS 225 Lab Report Week 4- Gram Staining

Examen ·

(3)

Biologie examenopgaven - Erfelijkheid

Examen ·

(0)

WGU C255 Terms Latest 2022 Rated A

Examen ·

(0)

NURS MISC Fundamentals Success 3e/SCORE A+

Examen ·

(0)

ATI Proctored Assessment Leadership 2019

Notes de cours ·

(0)

Samenvatting doelgroep volwassenen

Presentation ·

(0)

Presentatie AVG Hbo Rechten Jaar 1

Examen ·

(0)

NSG 5002 FINAL EXAM – QUESTION WITH ANSWERS

Examen ·

(0)

Enfermedades del aparato digestivo

$7.54
  • (0)
Ajouter au panier
Ajouté