LIFE202: Cell Signalling in Health & Disease

Topic 14: Life Decisions – Growth & Proliferation

KEY POINTS:
• The cell cycle is made up of 4 phases: G1 (gap 1); S phase, when the DNA is replicated; G2 (gap 2); and M
phase - replicated chromosomes separate (mitosis) & cell division takes place.
• Stages of the cell cycle are controlled by the cyclin-dependent kinases (CDKs) which are activated by proteins
known as the cyclins.
• Cyclin levels rise and fall through the cell cycle, with distinct cyclins associated with each phase.
• Specific cyclin–CDK complexes phosphorylate phase-specific targets, leading to the execution of that phase’s
program. These specific CDK complexes also control the timing of the transition to the next phase in the
cycle.
• CDK complexes initiate many feedback loops which play a central role in generating sharp and committed
transitions between phases of the cell cycle.
• Some key steps in the cell cycle are regulated by ubiquitin-mediated proteolysis, which makes these
transitions irreversible and committed.
• Cell-cycle progression can be arrested by checkpoint programs that detect key problems such as DNA
damage. These checkpoints provide time for the cell to fix problems before progressing with the cycle.

The Cell Cycle

Mitosis:
• Genome halves
• Cell division

Gap 2: Preparation for mitosis: Gap 0: ‘Quiescence’

• Protein synthesis or resting
• Rapid cell growth
• Reorganization of
microtubules

Gap 1:
• Cells grow in size
R Point • Increase supply of proteins
• Increase number of organelles
Synthesis: • Sense extracellular environment
DNA replicated • Checks DNA integrity

Weinberg, the Biology of Cancer (1st Edition, 2007)

In G1 phase, a cell has three options:

• To continue the cell cycle and enter S phase
• Exit the cell cycle and enter G0
• Become arrested in G1 phase (it may then enter G0 phase or re-enter cell cycle).
The deciding point is called the restriction (R) point or START and is regulated by proteins called
G1/S cyclins, which cause transition from G 1 to S phase. Passage through the G1 check point
commits the cell to division.
Growth factors provide environmental signals telling cells to divide

It’s important that cells only divide when appropriate. Growth

factors give cells the signal that they should start cell division.

Period during which

cells are responsive to
growth factors

Growth factors bind to cell surface receptors (receptor tyrosine kinases – see Topic 2) and induce
a signalling cascade. This results in the transcription of cyclins (specifically cyclin D) which cause
the transition from G1 to S phase:

Extracellular
growth factor

Growth factor
receptor

(cyclin D)

Add growth
factor

Cyclin D mRNA

Northern Blot (shows mRNA levels)

What are cyclins and how do they work?

cyclin D

Weinberg, the Biology of Cancer (1st Edition, 2007)

• Cyclins are proteins that interact with and activate cyclin-dependent kinases (CDKs)
• Cyclin levels rise and fall through the cell cycle, with distinct cyclins associated with each phase
• Cycles of transcription and proteolysis of each cyclin activates and inactivates CDKs at the correct time
• Specific cyclin–CDK complexes phosphorylate phase-specific targets that drive the cell cycle forward

Cyclins activate cyclin-dependent kinases (CDKs)

Cyclin D/E

Cyclin A

Cyclin B

Specific cyclins interact with specific CDKs. Cyclin binding to CDK has two effects:
1. Cyclins act as allosteric activators to shift the CDK into an active conformation;
2. Cyclins act as adaptors that bind to specific substrates, thus determining which proteins
CDK phosphorylates for a given phase of the cell cycle.

Each cyclin directs CDK to phosphorylate a distinct set of substrates that form the
physiological ‘program’ of that particular phase of the cell cycle.
How does expression of cyclin D switch on the cell cycle?

cyclin D

CDK1

• During G1, growth factors signal to increase

the expression of cyclin D (see above)
• Cyclin D interacts with and activates CDK4/6
• Active CDK4/6 phosphorylates retinoblastoma
protein Rb (a tumour suppressor protein)
CDK1 • Hyper-phosphorylated Rb releases E2F
transcription factors which can then induce the
expression of proteins (such as cyclin E) that
drive the transition from G1 to S phase (see
below).

Retinoblastoma protein Rb binds and inactivates E2F transcription factors until Rb is hyper-
phosphorylated by CDK4/6. Free and active E2Fs then drive G1/S transition

Activated
CyclinD/Cdk4/6

• E2F transcription factors induce the transcription of specific genes leading to the expression of
proteins that drive S phase eg Cyclin E
• Cyclin E/CDK2 then regulates DNA replication (S phase)
 Feedback regulation
In addition to executing the current phase program, CDK phosphorylation also leads to feedback
regulation that can help end the current cycle phase and drive entry into the next cycle phase.

Cyclins are not the only way to regulate CDK activity

• CDK inhibitors: CDK–cyclin complexes can

be inhibited by the binding of specific CDK
inhibitor subunits. Destruction of inhibitors
can be critical for activating CDKs.
• Activation of CDK also requires
phosphorylation in the activation loop of
the kinase
• Inhibitory phosphorylation site: CDK
activity can also be inhibited by
phosphorylation

Cyclin Dependent Kinase Inhibitors (CDKIs)

• Two major CDKI families in animal cells:

INK4 and CIP/KIP (both classified as
tumour suppressors)
• 90% of human cancers have abnormal
Rb, p16 or D-CDK4/6

CDK1 CDK1

Weinberg, the Biology of Cancer (1st Edition, 2007)

Destruction of cyclins turns off CDK activity

cyclin D

Decrease in
CDK1 activity

of cyclin B

• For example, during M phase, active CDK1/cyclin B phosphorylates Anaphase Promoting Complex
(APC, a ubiquitin E3 ligase)
• Phosphorylation of APC allows binding of CDC20
• CDC20 activates APC and also allows interaction with APC substrates (cyclin B)
• Leads to ubiquitination and proteolytic degradation of cyclin B and hence a decrease in CDK1
activity. Helps cycle move through the stages of mitosis (metaphase when the paired sister
chromatids align, to anaphase, when the sister chromatids are pulled apart by the mitotic
spindle)

The Cell Cycle is regulated and unidirectional due

to ‘checkpoints’

• Cell-cycle progression can be arrested by checkpoint

programs that detect key problems such as DNA
damage. These checkpoints provide time for the cell
to fix problems before progressing with the cycle.