Chapter 19 – The Eukaryotic Cell Cycle

Chapter 19 – The Eukaryotic Cell Cycle

19.1 Overview of the Cell Cycle

19.2 Model Organisms and Methods of Studying the Cell Cycle
19.3 Cell Cycle Progression and Control: Feedback Loops and
Posttranslational Modification
19.4 The Transition from G1 into S Phase and DNA Replication
19.5 The G2/M Transition and the Irreversible Engine of Mitosis
19.6 The Mitotic Spindle, Chromosome Segregation, and Exit from
Mitosis
19.7 Surveillance Mechanisms in Cell Cycle Regulation
19.8 Meiosis: A Special Type of Cell Division
The eukaryotic cell cycle
Chromosome duplication in S
Mitosis Stages
19.1 Overview of the Cell Cycle and Its Control
• Cell cycle 4 phases: G1, S, G2, and M
• Cells commit to division at the G1 START/Restriction point
• Cyclin-CDK complexes drive cell cycle progression
• Positive and negative feedback loops drive CDK activity
oscillations.
• Checkpoint pathway surveillance mechanisms guarantee
each cell cycle step is completed correctly before the next is
initiated.
 Chapter 19 – The Eukaryotic Cell Cycle

19.1 Overview of the Cell Cycle

19.2 Model Organisms and Methods of Studying the Cell Cycle
19.3 Cell Cycle Progression and Control: Feedback Loops and
Posttranslational Modification
19.4 The Transition from G1 into S Phase and DNA Replication
19.5 The G2/M Transition and the Irreversible Engine of Mitosis
19.6 The Mitotic Spindle, Chromosome Segregation, and Exit from
Mitosis
19.7 Surveillance Mechanisms in Cell Cycle Regulation
19.8 Meiosis: A Special Type of Cell Division
Human Cells undergoing Mitosis
DNA content by Flow
 The Eukaryotic Cell Cycle

19.2 Model Organisms and Methods of Studying the Cell Cycle

• Budding and fission yeast cell cycle mutants have revealed key
regulatory factors.
• Frog eggs and early embryos extracts have been used for
biochemical characterization of oscillatory cyclin-CDK complexes.
• Fruit flies provide a powerful system for investigating interplay
between cell division and developmental programs.
• Human tissue culture cells are used to study the properties of
the mammalian cell cycle.
 Chapter 19 – The Eukaryotic Cell Cycle

19.1 Overview of the Cell Cycle

19.2 Model Organisms and Methods of Studying the Cell Cycle
19.3 Cell Cycle Progression and Control: Feedback Loops and
Posttranslational Modification
19.4 The Transition from G1 into S Phase and DNA Replication
19.5 The G2/M Transition and the Irreversible Engine of Mitosis
19.6 The Mitotic Spindle, Chromosome Segregation, and Exit from
Mitosis
19.7 Surveillance Mechanisms in Cell Cycle Regulation
19.8 Meiosis: A Special Type of Cell Division
Network of CDKs
CDKs regulate cell cycle progression
Cyclin Binding
How Cyclins Were Discovered
CDK2 structure
 Cyclin-CDK activity
TABLE 19-1: Regulators of Cyclin-CDK Activity
Type of Regulator Function

Kinases and Phosphatases

CAK kinase Activates CDKs in all organisms

Wee1 kinase Inhibits CDKs in all organisms

Cdc25 phosphatase Activates CDKs in fission yeast

Cdcl4 phosphatase Activates Cdh1 to degrade mitotic cyclins in budding yeast

Cdc25A phosphatase Activates vertebrate S phase CDKs

Cdc25B and C phosphatases Activate vertebrate mitotic CDKs

Inhibitory Proteins

Sic1 Binds and inhibits S phase CDKs in budding yeast

CKIs p27KIP1, p57KIP2, and p21CIP Bind and inhibit CDKs in animal cells

INK4 Binds and inhibits G1 CDKs in animal cells

Rb Binds E2Fs, preventing transcription of multiple cell cycle genes in animal cells

Ubiquitin-Protein Ligases

SCF Degradation of phosphorylated Sic1 or p27KIP1 to activate S phase CDKs

APC/C Cdc20

Degradation of securin, initiating anaphase. Induces degradation of B-type cyclins

APC/C Cdh1

Degradation of B-type cyclins in G1 and geminin in metazoans to allow loading of replicative helicases on DNA replication origins

Phosphoserine/Threonine-Binding Proteins and Modular Domains

WD40 domains, leucine-rich repeats Recognize phosphorylated substrates and target them for degradation by SCF ligases

Cks subunits Target cyclin-CDKs for processive substrate phosphorylation

14-3-3 proteins
Bind and inactivate regulators of cyclin-CDKs to establish cell cycle checkpoints. Also regulates cytokinesis.

Pin1 Isomerizes phosphorylated substrates of CDKs to control mitotic entry and exit

FHA and BRCT domains

Form phospho-dependent molecular assemblies necessary for cell cycle surveillance mechanisms in all stages of the cell cycle, and regulates cytokinesis.

Polo-box domains Target Polo kinases to sites of CDK phosphorylation to drive mitotic entry and progression
CDK Subunit phosphorylation
CDK mutants
 The Eukaryotic Cell Cycle
19.3 Cell Cycle Progression and Control: Feedback Loops and Posttranslational
Modification
• Different cyclins present only in the cell cycle stage they promote activate
CDKs at different cell cycle stages.
• The ubiquitin-proteasome system limits presence of a cyclin to the
appropriate cell cycle stage.
• Activating and inhibitory phosphorylation of the CDK subunit regulates
CDK activity.
• CDK inhibitors (CKIs) inhibit CDK activity by binding directly to the cyclin-
CDK complex.
• CDKs initiate every aspect of each cell cycle stage by phosphorylating
many different target proteins.
• Phosphoserine/Threonine-Binding Domains Build Feedback Loops That
Coordinate CDK Activation and Cell Cycle Progression
 Chapter 19 – The Eukaryotic Cell Cycle

19.1 Overview of the Cell Cycle

19.2 Model Organisms and Methods of Studying the Cell Cycle
19.3 Cell Cycle Progression and Control: Feedback Loops and
Posttranslational Modification
19.4 The Transition from G1 into S Phase and DNA Replication
19.5 The G2/M Transition and the Irreversible Engine of Mitosis
19.6 The Mitotic Spindle, Chromosome Segregation, and Exit from
Mitosis
19.7 Surveillance Mechanisms in Cell Cycle Regulation
19.8 Meiosis: A Special Type of Cell Division
 The Eukaryotic Cell Cycle (3 of 7)

19.4 The Transition from G1 into S Phase and DNA Replication

• Extracellular signals − nutritional state (in yeast) and
presence of mitogens and anti-mitogens (in vertebrates) −
regulate cell cycle entry.
• Molecular events promoting entry into the cell cycle are
conserved across species.
• G1/S CDKs trigger chromosome duplication at DNA origin of
replication sites.
• Cohesins link replicated DNA molecules to ensure accurate
segregation during mitosis.
 Chapter 19 – The Eukaryotic Cell Cycle

19.1 Overview of the Cell Cycle

19.2 Model Organisms and Methods of Studying the Cell Cycle
19.3 Cell Cycle Progression and Control: Feedback Loops and
Posttranslational Modification
19.4 The Transition from G1 into S Phase and DNA Replication
19.5 The G2/M Transition and the Irreversible Engine of Mitosis
19.6 The Mitotic Spindle, Chromosome Segregation, and Exit from
Mitosis
19.7 Surveillance Mechanisms in Cell Cycle Regulation
19.8 Meiosis: A Special Type of Cell Division
Positive feedback loop
Nuclear envelope dissolution
Centrosome
Mitotic spindle attachment
 The Eukaryotic Cell Cycle (4 of 7)
19.5 The G2/M Transition and the Irreversible Engine of Mitosis
• Mitotic CDKs induce entry into mitosis in all eukaryotes by inducing
chromosome condensation, nuclear envelope breakdown, and
spindle formation.
• Mitotic CDKs:
– are inactivated by inhibitory phosphorylation of the CDK subunit until
completion of DNA replication.
– promote their own activation through positive feedback loops that
inactivate Wee1 kinase and activate Cdc25 phosphatase.
• The kinetochore on each compacted sister chromatid attaches to
microtubules emanating from opposite spindle poles.
• Cells ensure bi-orientation of sister chromatids in the spindle by a
tension-based mechanism.
 Chapter 19 – The Eukaryotic Cell Cycle

19.1 Overview of the Cell Cycle

19.2 Model Organisms and Methods of Studying the Cell Cycle
19.3 Cell Cycle Progression and Control: Feedback Loops and
Posttranslational Modification
19.4 The Transition from G1 into S Phase and DNA Replication
19.5 The G2/M Transition and the Irreversible Engine of Mitosis
19.6 The Mitotic Spindle, Chromosome Segregation, and Exit from
Mitosis
19.7 Surveillance Mechanisms in Cell Cycle Regulation
19.8 Meiosis: A Special Type of Cell Division
Separase regulation
Nuclear envelope reassembly
RhoA regulates cytokinesis
 The Eukaryotic Cell Cycle (5 of 7)

19.6 The Mitotic Spindle, Chromosome Segregation, and Exit

from Mitosis
• Cohesin cleavage by separase initiates chromosome
segregation during anaphase.
• Exit from mitosis is triggered by mitotic cyclin
degradation and requires protein phosphatase reversal of
mitotic CDK phosphorylation of many different proteins,
permitting mitotic spindle disassembly, decondensation
of chromosomes, and reassembly of the nuclear
envelope.
• Cytokinesis position is coordinated with spindle position.
 Chapter 19 – The Eukaryotic Cell Cycle

19.1 Overview of the Cell Cycle

19.2 Model Organisms and Methods of Studying the Cell Cycle
19.3 Cell Cycle Progression and Control: Feedback Loops and
Posttranslational Modification
19.4 The Transition from G1 into S Phase and DNA Replication
19.5 The G2/M Transition and the Irreversible Engine of Mitosis
19.6 The Mitotic Spindle, Chromosome Segregation, and Exit from
Mitosis
19.7 Surveillance Mechanisms in Cell Cycle Regulation
19.8 Meiosis: A Special Type of Cell Division
DNA damage response system
Cell cycle checkpoints
Spindle assembly checkpoint
 The Eukaryotic Cell Cycle (6 of 7)

19.7 Surveillance Mechanisms in Cell Cycle Regulation

• Checkpoint pathways ensure the next cell cycle event does
not occur prior to the completion of the preceding one.
• Checkpoint pathways are comprised of event sensors, a
signaling pathway, and an effector that halts cell cycle
progression and activates repair pathways when necessary.
• Checkpoint pathways monitor and respond to DNA
replication and damage, and spindle assembly and position.
• The highly conserved Hippo signal pathway coordinates fungi
chromosome segregation with mitosis exit and cytokinesis
but coordinates metazoan tissue growth with tissue
organization.
 Chapter 19 – The Eukaryotic Cell Cycle

19.1 Overview of the Cell Cycle

19.2 Model Organisms and Methods of Studying the Cell Cycle
19.3 Cell Cycle Progression and Control: Feedback Loops and
Posttranslational Modification
19.4 The Transition from G1 into S Phase and DNA Replication
19.5 The G2/M Transition and the Irreversible Engine of Mitosis
19.6 The Mitotic Spindle, Chromosome Segregation, and Exit from
Mitosis
19.7 Surveillance Mechanisms in Cell Cycle Regulation
19.8 Meiosis: A Special Type of Cell Division
Meiosis
 The Eukaryotic Cell Cycle (7 of 7)

19.8 Meiosis: A Special Type of Cell Division

• Meiosis involves one cycle of chromosome replication
followed by two cycles of cell division to produce
haploid germ cells.
• Meiosis-specific gene products and activities modulate
the mitotic cell division program to perform meiosis.