2009 Lecture 15

From Cellbiology
Jump to: navigation, search

Contents

Cell Cycle

The Cell Cycle

Introduction

This lecture will introduce the cell cycle, which is the entire life of a cell from birth to death. This overall topic is too large for a single lecture, so will focus on our understanding of the regulation of the cell cycle. Later lectures will cover cell birth (cell division), differentiation (signaling, development) and death (2 lectures) in more detail.

Dr Mark Hill (2009)

Lecture Audio

The University has a system for automated recording of lectures called Lectopia. Lectopia requires login using your student number and unipass. I will be adding the link to each iLecture Audio following the Lecture. Due to the automated recording method, most lectures begin 4-5 minutes into MP3 recordings and occasionally stop before the end of the lecture. 2009 All Audio Files

Archive

MH - note that content listed below will not match exactly current lecture structure but has been selected as having similar content.

Objectives

  • Understanding of the main stages of the cell cycle
  • Broad understanding of different cell lifespan
  • Understanding of the differences between embryonic and adult cell cycles
  • Understanding of the key regulators of cell cycle
    • cyclins
    • cyclin dependent kinases
  • Broad understanding of abnormal cell cycles
    • How the cell cycle is monitored

Background Reading

Nature - Cell Division Milestones


Recent Nobel Prizes

2001 - Control of the Cell Cycle

  • Leland Hartwell (born 1939), Fred Hutchinson Cancer Research Center, Seattle, USA, is awarded for his discoveries of a specific class of genes that control the cell cycle. One of these genes called "start" was found to have a central role in controlling the first step of each cell cycle. Hartwell also introduced the concept "checkpoint", a valuable aid to understanding the cell cycle.
  • Paul Nurse (born 1949), Imperial Cancer Research Fund, London, identified, cloned and characterized with genetic and molecular methods, one of the key regulators of the cell cycle, CDK (cyclin dependent kinase). He showed that the function of CDK was highly conserved during evolution. CDK drives the cell through the cell cycle by chemical modification (phosphorylation) of other proteins.
  • Timothy Hunt (born 1943), Imperial Cancer Research Fund, London, is awarded for his discovery of cyclins, proteins that regulate the CDK function. He showed that cyclins are degraded periodically at each cell division, a mechanism proved to be of general importance for cell cycle control.

1989 - Cellular Origin of Retroviral Oncogenes

  • Michael Bishop and Harold Varmus used an oncogenic retrovirus to identify the growth-controlling oncogenes in normal cells. In 1976 they published the remarkable conclusion that the oncogene in the virus did not represent a true viral gene but instead was a normal cellular gene, which the virus had acquired during replication in the host cell and thereafter carried along.

Prokaryote Division

  • Binary Fission
    • and seen in eukaryote mitochondria
  • Asexual reproduction
  • replicates original cell to produce two identical cells
  • Grow in numbers exponentially
  • adequate nutrients and a fast life cycle
  • single organism can multiply into billions
  • High mutation rate of bacteria

Appear to involve proteins that are homologs of eukaryotic cytoskeleton proteins. Prokaryotic Cytoskeleton Filaments

Cell Lifespan

  • Body Cell Types - about 210 types
  • Lifespan
    • Born
    • Differentiate
    • Function
    • Die

Cell Lifespan Examples

Neutrophil 6-7 hours circulating 4 days in tissue Red blood cell 120 days Brain neuron, heart 50 - 100 years

Lifespan Processes

  • Birth

Mitosis Except germ cells Meiosis

  • Death

Apoptosis Programmed cell death Necrosis

Movie: Embryo Mitosis Movie: Cell Death (Apoptosis)

Cell Cycle Major Phases

  • Mitosis (M phase)

Cell birth(division) small time of cell cycle

  • Interphase

Most cell life Cell growth, function DNA synthesis organelle development

Cell Cycle Time cell comes into existence until that cell divides again Rapidly growing human cells 20-24 hr Liver cells 1-2 year Neurons 1 only Quiescent G0

Cell Cycle- Stages

Rapidly dividing cell (20-24hr)

Mitosis

  • M phase 1 hr

Interphase

  • G1 Phase
    • cellular growth 9hr
    • Most variable time
    • Can exit to G0
  • S Phase
    • DNA duplication 9hr
  • G2 Phase
    • growth prepare for mitosis 4 hr

Cell Cycle Differences

Early embryonic cycle

  • no growth occurs
  • each daughter cell is half the size of parent cell
  • cycle time is very short
  • S phases and M phases alternate without any intervening G1 or G2 phases

Cell Cycle Regulation

Cell proliferation is strictly regulated

Unregulated/abnormal proliferation is oncogenesis or Cancer

Cell-Cycle Control


An overview of the cell-cycle control system

Cell Cycle- External Regulators

  • Cell replacement in different tissues
    • regulated by growth factors
    • can be specific for specific cell types

Growth Factors and Cell Cycle Progress

  • External factors can also regulate progression through cycle
  • Growth factors primarily act on cells in G0 and G1
  • The restriction point is the timepoint in G1 when cells no longer respond to withdrawal of growth factors by returning to G0, but progress to S phase.
    • thought to involve retinoblastoma protein (pRb)

Restriction Point

Growth Factor Model

  • Fibroblasts in culture
    • Serum (Prepared by clotting)- Proliferation
    • Plasma (Prepared by centrifugation, no clotting)- no proliferation
  • Clotting
    • allows platelets to release secretory granules
    • Platelet-derived growth factor (PDGF)
  • Connective tissue cells express PDGF receptors which bind the small PDGF glycoprotein

Other Growth Factors

  • Interleukin-2 (IL-2)
    • Stimulates T lymphocytes
  • Nerve Growth Factor (NGF)
    • Promotes neuronal survival and growth
  • Epidermal Growth Factor (EGF)
  • Vascular Endothelial Growth Factor (VEGF)
  • Insulin-like growth factors (IFG-1, IGF-2)

Cell Cycle- Internal Regulators

1980s - studies in Xenopus eggs and starfish oocytes purified M phase-promoting factor (MPF) from and the identification of its components as cyclin B and CDC2 (also called cyclin-dependent kinase

Cyclins and Cyclin-dependent Kinases

Cyclins

Cyclins are synthesized and degraded each cell cycle (hence the name)

Cyclins and Cyclin-Dependent Kinases need to interact for cell cycle progression Cyclins and Cyclin-Dependent Kinases

Cell cycle1.jpg

Cyclin D

Cyclin D Interactions
  • cyclin D1, D2 and D3
  • expression induced by growth factors stimulation
    • serum growth factors to quiescent cells promotes transcription of the cyclin D1 gene
  • Cyclin D1 then binds Cdk4 and Cdk6 in early to mid-G1 phase
    • phosphorylate and inactivate retinoblastoma protein (pRb)
  • also acts as a cofactor for several transcription factors in numerous cell types

Cyclin D1 is a proto-oncogene

  • mutations are associated with cancer progression
  • as a regulator of G1 to S-phase transition

Cyclin E

  • required for transition from G1 to S phase
  • Cyclin E binds to G1 phase Cdk2
  • Cyclin E/Cdk2 complex phosphorylates p27Kip1 which then degrades
    • an inhibitor of Cyclin D
  • activation of Cyclin E gene can be blocked by the cdk inhibitor p16 (Cyclin-dependent kinase inhibitor 2A)
    • tumor suppressor protein
  • expression of Cyclin A then increases for progress to S phase

Cyclin A

  • accumulates from early S phase
    • role not fully understood, required for S phase progress
  • Cyclin A binds Cdk2
  • disappears ahead of cyclin B during mitosis
  • can bind to both Cdc2 and Cdk2

Cyclin B

  • accumulates from S phase
  • Cyclin B forms a complex with Cdc2
    • complex is kept inactive by phosphorylation of Cdc2
    • abruptly activated by Cdc25 during mitosis
  • cyclin B is destroyed at mitosis exit by ubiquitin-mediated mechanism (catalyzed by the APC/C)

control of proteolysis by SCF and APC during the cell cycle

Cyclin-dependent Kinases

Inactive until bound to a specific cyclin

kinase - means an enzyme which phosphorylates a target protein(s)

  • Drive M to S Phase
    • cdk1 and cdk2
  • Cdk1 activated at G2 to M
  • Cdk2 activated at G1 to S

Cell Cyclins - Interphase Cell Cyclins - M Phase

  • Division controlled by synthesis/degradation cyclin B
  • regulatory subunit of Cdc2 protein kinase
  • interphase cyclin B synthesis leads to formation of active cyclin B–Cdc2 complex
  • induces entry into mitosis

Rapid degradation of cyclin B leads to inactivation Cdc2 kinase

  • Allow cell to exit mitosis and return to interphase next cell cycle


  • cyclin B-CDC2 acts as M phase-promoting factor (MPF)
    • activate other proteins through phosphorylation
  • cyclin A down-regulation induces a G2 phase arrest through a checkpoint-independent inactivation of cyclin B-CDC2 by inhibitory phosphorylation.
    • cyclin A cannot form MPF independent of cyclin B

Regulator Checkpoints

These regulators are often described as "tumor suppressor proteins" due to their ability to block tumor (cancer) growth. Conversely, mutations in these genes often lead to tumor growth.

Checkpoints in the cell-cycle control system

How DNA damage arrests the cell cycle in G1

p53

p53 pathways
  • (TP53) A multifunctional protein Mr 53 kDa regulating cell cycle and apoptosis
  • As a cell cycle regulator it recognizes and binds damaged DNA
    • single-stranded DNA, insertion/deletion mismatches, and free DNA ends
  • Acts as a transcription factor activating p21 transcription
    • p21 protein then inhibits G1 cyclin-dependent kinases p21

p53-induced cell-cycle arrest in response to DNA damage

unstressed cells

  • p53 levels are low through a continuous degradation
  • Mdm2 (HDM2 in humans)
    • binds p53
    • blocks action
    • transports p53 from nucleus to the cytosol
  • gene has been shown to be damaged in cells by mutagens
  • mutations found with

Effect of mutation of the p53 tumorsuppressor gene on G1 DNA-damage checkpoint control

Science magazine designated p53 the 'Molecule of the Year' for 1993

pRb

pRb Interactions
  • retinoblastoma protein regulating cell cycle
  • As a cell cycle regulator it recognizes damaged DNA
  • binds and inhibits transcription factors of the E2F family
    • active pRb is hypophosphorylated
    • inactive pRb is phosphorylated
  • mutations in this gene lead to the disease retinoblastoma OMIM 180200

p16

Restriction point control

  • also called Cyclin-dependent kinase inhibitor 2A
  • blocks activation of Cyclin E gene
  • mutations found with
    • pancreatic adenocarcinoma OMIM 260350
    • esophageal and gastric cancer cell lines

Oncogenesis

Cancer requires multiple mutations

MH - There are many detailed resources on oncogenesis (cancer) this is only a brief mention in regard to cell cycle. See also the tumor suppressor section above. An importanrt issue is the concept of "multiple hits".

  • some viruses can infect human cells and lead to oncogenesis (cervical cancer, liver cancer, and certain lymphomas, leukemias, and sarcomas)
    • for example - human papillomavirus (HPV)

References

Textbooks

Essential Cell Biology

  • Essential Cell Biology Chapter 17

Molecular Biology of the Cell

Alberts, Bruce; Johnson, Alexander; Lewis, Julian; Raff, Martin; Roberts, Keith; Walter, Peter New York and London: Garland Science; c2002

Molecular Cell Biology

Lodish, Harvey; Berk, Arnold; Zipursky, S. Lawrence; Matsudaira, Paul; Baltimore, David; Darnell, James E. New York: W. H. Freeman & Co.; c1999

The Cell- A Molecular Approach

Cooper, Geoffrey M. Sunderland (MA): Sinauer Associates, Inc.; c2000

Search Online Textbooks

Books

PubMed

  • PubMed is a service of the U.S. National Library of Medicine that includes over 18 million citations from MEDLINE and other life science journals for biomedical articles back to 1948. PubMed includes links to full text articles and other related resources. PubMed
  • PubMed Central (PMC) is a free digital archive of biomedical and life sciences journal literature at the U.S. National Institutes of Health (NIH) in the National Library of Medicine (NLM) allowing all users free access to the material in PubMed Central. PMC
  • Online Mendelian Inheritance in Man (OMIM) is a comprehensive compendium of human genes and genetic phenotypes. The full-text, referenced overviews in OMIM contain information on all known mendelian disorders and over 12,000 genes. OMIM
  • Entrez is the integrated, text-based search and retrieval system used at NCBI for the major databases, including PubMed, Nucleotide and Protein Sequences, Protein Structures, Complete Genomes, Taxonomy, and others Entrez

Search Pubmed

Reviews

  • The great divide: coordinating cell cycle events during bacterial growth and division. Haeusser DP, Levin PA. Curr Opin Microbiol. 2008 Apr;11(2):94-9. Epub 2008 Apr 7. Review. PMID: 18396093 | PMC
  • Cell cycle studies based upon quantitative image analysis. Stacey DW, Hitomi M. Cytometry A. 2008 Apr;73(4):270-8. Review. PMID: 18163464
  • Analysis of cell cycle phases and progression in cultured mammalian cells. Schorl C, Sedivy JM. Methods. 2007 Feb;41(2):143-50. Review. PMID: 17189856
  • Cell cycle regulation of DNA replication. Sclafani RA, Holzen TM. Annu Rev Genet. 2007;41:237-80. Review. PMID: 17630848


OMIM

Links

2009 Course Content

Lectures

Cell Biology Introduction | Cells Eukaryotes and Prokaryotes | Cell Membranes and Compartments | Cell Nucleus | Cell Export - Exocytosis | Cell Import - Endocytosis | Cell Mitochondria | Cell Junctions | Cytoskeleton Introduction | Cytoskeleton 1 Intermediate Filaments | Cytoskeleton 2 Microtubules | Cytoskeleton 3 Microfilaments | Extracellular Matrix 1 | Extracellular Matrix 2 | Cell Cycle | Cell Division | Cell Death 1 | Cell Death 2 | Signal 1 | Signal 2 | Stem Cells | Stem Cells | Development | Revision

Laboratories

Introduction to Lab | Microscopy Methods | Preparation/Fixation | Immunochemistry | Cell Knockout Methods | Cytoskeleton Exercise | Confocal Microscopy | Tissue Culture 1 | Tissue Culture 2 | Microarray Lab visit

Dr Mark Hill 2013, UNSW Cell Biology - UNSW CRICOS Provider Code No. 00098G