The Cell Cycle

Molecular Biology of the Cell

An Overview of the Cell Cycle

1. The eucaryotic cell cycle is divided into four phases

2. Cell-cycle control is similar in all eucaryotes
3. Cell-cycle control system dissected genetically in yeasts
4. Cell-cycle control system analyzed biochemically in animal embryos
5. Cell-cycle progression studied in various ways

The major events of the cell cycle

Events of eucaryotic cell division as seen under a microscope

The four phases of the cell cycle

Cell-cycle control system dissected genetically in yeasts

Behavior of a temperature-sensitive Cdc mutant

room temperature

36C

Morphology of budding yeast cells arrested by a Cdc mutation

Normal yeast cells buds vary in size

according to the cell-cycle stage

In a Cdc15 mutant, grown at the

restrictive temperature, cells complete
anaphase but cannot complete the exit
from mitosis and cytokinesis. They
arrest uniformly with the large buds,
which are characteristic of late M phase

Cell-cycle control system analyzed biochemically in animal embryos

A mature Xenopus egg, ready for fertilization

Oocyte growth and egg cleavage in Xenopus

Cell-cycle progression studied in various ways

Labeling S-phase cells

Analysis of DNA content with a flow cytometer

The cell-cycle control system

1.

Cell-cycle control system triggers the major events of the cell cycle

2.

The cell-cycle control system depends on cyclically activated cyclindependent protein kinases (Cdks)

3.

Inhibitory phosphorylation and Cdk inhibitory proteins (CKIs) can

suppress Cdk activity

4.

The cell-cycle control system depends on cyclical proteolysis

5.

Cell-cycle control also depends on transcriptional regulation

6.

The cell-cycle control system functions as a network of biochemical

switches

Control of the cell cycle

Two key components of the cell-cycle control system

Cyclin-Cdk complexes of the cell-cycle control system

Inhibitory phosphorylation and Cdk inhibitory proteins (CKIs)

can suppress Cdk activity

The structural basis of Cdk activation

The regulation of Cdk activity by inhibitory phosphorylation

The inhibition of a cyclin-Cdk complex by a CKI

The cell-cycle control system depends on cyclical proteolysis

Cell-cycle control also depends on transcriptional regulation

In budding yeast, about 10% of the genes encode mRNAs

whose levels oscillate during the cell cycle

The cell-cycle control system functions as a network of

biochemical switches

An overview of the cell-cycle control system

The two central events of the cell cycle are:

- replication of DNA during the S phase
- chromosome segregation and cell division during the M phase
Both these events are controlled by the cyclin-Cdk complexes

S phase

1.

S-Cdk initiates DNA replication once per cycle

2.

Chromosome duplication requires duplication of chromatin structure

3.

Cohesins help hold sister chromatids together

Control of chromosome duplication

Control of the initiation of DNA replication

The ORC remains associated with the ori site
throughout the cell cycle.
In early G1, Cdc6 and Cdt1 (helicase loading
proteins) associate with the ORC and the
resulting complex allows the assembly of the
Mcm ring and the formation of the
prereplicative complex.
In the S phase, S-Cdk stimulates the assembly
of several additional proteins to form the
preinitiation complex. Other proteins are
recruited to the origin and replication begins.
S-Cdk blocks rereplication by triggering the
destruction of Cdc6 and the inactivation of
the ORC.
The cell is able to assemble the pre-RC only
after M-Cdk is inactivated and APC/C is
activated at the end of the M-phase

S-Cdk activity is high during G2 and early mitosis. This prevents

rereplication from occurring after the S phase
M-Cdk also prevents rereplication from occurring during mitosis by
phosphorylating the Cdc6 and ORC proteins
With all the control elements preventing rereplication,
how does DNA replication take place in the next cell
cycle?
At the end of mitosis, APC/C activation leads to the inactivation of
Cdk activity and the destruction of geminin. Pre-RC components are
dephosphorylated and Cdt1 is activated allowing pre-RC assembly
to initiate a new round of replication

Mitosis
1. M-Cdk drives entry into mitosis
2. Dephosphorylation activates M-Cdk at the onset of
mitosis
3. Condensin helps configure duplicated chromosomes for
separation
4. The mitotic spindle is a microtubule-based machine
5. Centrosome duplication occurs early in the cell cycle
6. M-Cdk initiates spindle assembly in prophase
7. The completion of spindle assembly in animal cells
requires nuclear envelope breakdown
8. The APC/C triggers sister-chromatid separation and the
completion of mitosis
9. Unattached chromosomes block sister-chromatid
separation: The spindle assembly checkpoint

Activation of M-Cdk drives entry into mitosis

The APC/C triggers sister-chromatid separation and the

completion of mitosis

Control of cell division and cell growth

Mechanism controlling
cell-cycle entry and
S-phase initiation
in animal cells

How DNA damage

arrests the cell cycle
in G1