The Cell Cycle

The Cell Doctrine – Rudolf Virchow 1858

 
 
“Where a cell arises, there must be a previous
cell, just as animals can only arise from animals
and plants from plants”
 
• Cells are generated from cells and the only way
to make more cells is by the division of those
that already exist.

• All living organisms, from the unicellular

bacterium to the multicellular mammal, are
products of repeated rounds of growth and
division extending back in time to the
beginnings of life on Earth over three billion
years ago.
The Events of Eukaryotic Cell Division as Seen
Under a Microscope
• Eukaryotic cells have evolved a complex network of
regulatory proteins, known as the cell cycle control system,
which governs the progression through the cell cycle.

• The core of this system is an ordered series of biochemical

switches that control the main events of the cycle, including
DNA replication and the segregation of the replicated
chromosomes.

• In most cells, additional layers of regulation enhance the

fidelity of cell division and allow the control system to
respond to various signals both outside and inside the cell.
• Inside the cell, the control system monitors
progression through the cell cycle and delays later
events until earlier events have been completed.

• The cell-cycle control system therefore has a

central role in regulating cell numbers in the
tissues of the body. When the system
malfunctions excessive cell divisions can result in
cancer.
The Phases of the Cell Cycle
 G phases
• The two gap phases serve as more than simple time
delays to allow cell growth. They also provide time
for the cell to monitor the internal and external
environment to ensure that conditions are suitable
and preparations are complete before the cell
commits itself to S and M phases of the cycle.

• The G1 phase is especially important in this respect.

Its length can vary greatly depending on external
conditions and extracellular signals from other cells.
 G phases
• If extracellular conditions are unfavorable, for
example, cells delay progress through G1 and
may even enter a specialized resting state,
known as G0, in which they can remain for days,
weeks or even years before resuming
proliferation.

• Many cells (e.g. neurons) remain permanently in

G0 until they or the organism dies.
 G phases
• If extracellular conditions are favorable and signals
to grow and divide are present, cells in early G1 or
G0 progress through a commitment point near the
end of G1, termed start (in yeasts) or the restriction
point (in mammals).

• After passing this point cells are committed to DNA

replication even if the extracellular signals that
stimulate cell growth and division are removed.
 The cell cycle control system should and does possess the following
features

• A clock, or timer, that turns on each event at a specific

time, thus providing a fixed amount of time for the
completion of each event.

• A mechanism for initiating events in the correct order;

entry into mitosis must always come after DNA
replication

• A mechanism to insure that each event is triggered

only once per cycle.
 The cell cycle control system should and
does possess the following features
• Binary (on/off) switches that trigger events in a complete
irreversible fashion. It would clearly be disastrous if events
were initiated but not completed.

• Robustness: backup mechanisms to ensure that the cycle

can work properly even when parts of the system
malfunction

• Adaptability so that the systems behaviour can be modified

to suit specific cell types or environmental conditions.
• Progression through G1 and G2 is delayed by breaking
mechanisms if the DNA in the chromosomes is damaged by
radiation or chemicals.

• Delays at these DNA damage checkpoints provide time for the

damaged DNA to be repaired, after which the cell cycle brakes
are released and progress resumes.

• Checkpoints are important as they also act as points in the cell

cycle at which the control system can be regulated by
extracellular signals from other cells.

• These signals – that can either promote or inhibit cell

proliferation – tend to act by regulating progression through a G1
checkpoint.
Checkpoints in the Cell Cycle Control System
 Checkpoint Mechanisms
• Tend to act through negative intracellular signals that
arrest the cell cycle, rather than through the removal of
positive signals that normally stimulate cell-cycle
progression.

• For example, in a cell with many chromosomes, if each

chromosome set a positive signal to the cell cycle control
mechanism once it’s attached to the spindle, the
attachment of the last chromosome would be hard to
detect, as it will be signaled by only a small fractional
change in the total go signal.
 Checkpoint Mechanisms
• If on the other hand each unattached chromosome sends
a negative signal to inhibit progress through the cell cycle,
the attachment of the last chromosome will be easily
detected because it will cause a change from some stop
signal to none.

• Although most checkpoints are not essential for cell cycle

progression under ideal conditions, checkpoints are best
viewed as accessory braking systems that have been
added to the cell cycle control system to provide a more
sophisticated form of regulation.
Molecular Control of the Cell cycle
• Cyclin dependent
Kinases (Cdks) are the
central components of
the cell cycle. They are
inactive unless a cyclin
is bound
A Simplified View of the Core of the Cell
Cycle Control System
• Rising G1/S cyclins trigger progression through start

• S-Cdk trigger DNA replication as well as some early mitotic events

• M-Cdk complexes form during G2 are held in an inactive state. Activated at the end of
G2 and trigger early events of mitosis
 Cdk Proteins
• In yeast cells, a single Cdk protein binds all
classes of cyclins and drives all cell cycle
events by changing cyclin partners at different
stages of the cycle.

• In contrast, in vertebrate cells there are four

Cdks. Two interact with G1-cyclins, one with
G1/S and S-cyclins, and one with M-cyclins.
 Cyclins
• There are 4 classes of cyclins, each defined by the stage of the
cell cycle at which they bind Cdks and function.
• Three of these classes are required in all eukaryotic cells

• G1/S-cyclins bind Cdks at the end of G1 and commit the cell to

DNA replication
• S-cyclins bind Cdks during S phase are required for the initiation
of DNA replication.
• M-cyclins promote the events of mitosis.
 
• In most cells a fourth class of cyclins, the G1-cyclins helps to
promote passage through the start or restriction point in late G1.
 How do different cyclin-Cdk complexes
drive different cell cycle events?
• The answer in part appears to that the cyclin protein not only
activates its partner, but also directs it to specific target
proteins.

• As a result each cyclin-Cdk complex phosphorylates a different

set of substrate proteins.

• The same cyclin-Cdk complex can also induce different effects

at different times in the cycle, probably because the
accessibility of some Cdk substrates changes during the cell
cycle e.g. certain proteins that function only in mitosis, may
become available for phosphorylation only in G2.
The Structural Basis of Cdk Activation
Additional Regulatory Mechanisms

This regulatory mechanism is particularly important

in the control of M-Cdk activity at the onset of
mitosis
 Additional Regulatory Mechanisms

There are a variety of CKI proteins and they are primarily employed in the
control of G1 and S phase.
Analysis of the three dimensional structure of a cyclin-Cdk-CKI complex
has revealed that CKI binding dramatically rearranges the structure of
the Cdk active site, rendering it inactive
 Cyclical Proteolysis

The rate limiting step in cyclin destruction is the

final ubiquitin-transfer reaction catalyzed by
enzymes known as ubiquitin ligases.
 Anaphase Promoting Complex
• In M phase, the Anaphase Promoting Complex
(APC) is responsible for the ubiquitylation and
proteolysis of M-and S-cyclins and other
regulators of mitosis. Destroying these cyclins
inactivates most Cdks in the cell

• It is the key regulator of the metaphase to

anaphase transition
APC
• As a result the many proteins phosphorylated
by Cdks from S phase to early mitosis are
dephosphorylated. The dephosphorylation of
Cdk targets is required for the completion of M
phase.

• Following its activation APC remains active in G1

providing a stable period of Cdk inactivity.

• When G1/S cyclins are activated in late G1 APC

is turned off
• APC also catalyses the ubiquitylation and
destruction of securin which normally protects
the protein linkages that hold sister chromotid
pairs together in early mitosis.

• This activates a protease that separates the

sisters an unleashes anaphase.
 SCF
• In G1 and S-phase, an enzyme complex termed
SCF is responsible for the ubiquitylation and
destruction of G1/S cyclins and certain CKI
proteins that control S phase initiation.

• Phosphorylated CKIs are recognized by substrate

binding subunits of the SCF complex called F-box
proteins of which there are many different types
SCF
 Regulation
• APC activity changes during the cell cycle in
response to its association with activating
proteins i.e. Cdc20 (anaphase) or CdhI (late
mitosis - early G1).

• SCF activity is constant during the cell cycle.

Ubiquitylation is controlled by changes in the
phosphorylation state of its target proteins
 Transcriptional Control
• Cyclin levels in most cells are controlled not
only by the changes in cyclin degradation but
also by changes in cyclin gene transcription and
cyclin synthesis.

• Use of DNA arrays of organisms such as budding

yeast allow analysis of changes in the
expression of all the genes in the genome as the
cell progresses through the cell cycle.
• From these experiments it has been demonstrated that about
10% of the yeast genes encode mRNAs whose levels oscillate
during the cell cycle.

• Some of these genes encode proteins with known functions in

cell cycle control, but the functions of many others are unknown.

• It is however likely that these oscillations in gene expression are

controlled by the cyclin-Cdk dependent phosphorylation of gene
regulatory proteins, but the details of this regulation remain
unknown.
Overview of the Cell Cycle Control System