Faculty member: Asst.Prof.

Süreyya Bozkurt
E-mail: sbozkurt@istinye.edu.tr
Department: Medical Biology
Lesson:Cell Cycle
 Learning Outcomes

• Learns the definition of cell cycle

• Learns the names and functions of proteins that function in the cell cycle
• Learns the control steps and mechanisms of the cell cycle
 - The main mechanism for the growth of all
living organisms and the formation of
identical cells is the duplication and division
of existing cells.

- Cycle of duplication and division, known

as the cell cycle

- Each cell division in unicellular

organisms (such as bacteria, yeast)
forms a new organism.

- It functions in multicellular organisms in

the formation of the new organism in the
embryonic stage, in replacing the dead cells
in the adult stage.
Molecular biology of the cell sixth edition (Bruce Alberts et.al)
The major events of the cell cycle:

1- Duplication of the genetic material (DNA) (S

Phase)

2- Segregation of the genetic material into

genetically identical daughter cells and cytokinesis
(M Phase)

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

 The events of eukaryotic cell division as seen under a microscope

The nuclear division (mitosis) and cell division (cytokinesis), collectively called M phase, typically
occupy only a small fraction of the cell cycle. The other, much longer, part of the cycle is known
as interphase
-One cell cycle of a typical cultured human cell requires 24 hours
-1 hour for M phase and 23 hours for interphase

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

 The four phases of the eukaryotic cell cycle

-Eukaryotic cell cycle consists of 4 sequential

phases

Gap1 (G1), Synthesis (S), Gap2 (G2) and Mitosis

(M) phases. G1 + S + G2 = Interphase

G (Gap) phases are phases in which the internal

and external conditions are appropriately
checked and preparations are completed before
entering into the S phase and M phase.

-G1 phase is particularly important, and the

length of this phase varies depending on the
external conditions and extracellular signals
Molecular biology of the cell sixth edition (Bruce from other cells.
Alberts et.al)
-If the conditions are not suitable, the cells enter the G0
phase and may remain in days, months or even years in
this phase. Some cells remain in the G0 phase until the
organism dies.

-Many cells in our body are in G0 phase (neurons, skeletal

muscle cells). Some cells enter and exit the cell cycle
when needed (exp. damaged liver cells).

-Some enter the cell cycle as repetitions (fibroblasts and

lymphocytes).
 Phases of the cell cycle can be observed by various methods

Microscopic observation Immunofluorescence

staining

Analysis of DNA content with flow cytometry

Molecular biology of the cell sixth edition
(Bruce Alberts et.al)
A cell-cycle control system triggers the essential processes of the cycle
The cell-cycle control system is based on a connected series of
biochemical switches, each of which initiates a specific cell-
cycle event such as DNA replication, mitosis, and cytokinesis.

-Most of the eukaryotic cells have 3 main control points:  

1-start transition (restriction point): It is seen in late stages of

G1. Evaluates the suitability of the environmental conditions
for the cell to enter the cell cycle.

2-G2/M transition: triggers early mitotic events. The cell checks

the environmental factors before entering the mitosis and
whether the DNA is replicated correctly.

3-Metaphase-to-anaphase transition: Checks whether all

chromosomes are correctly attached to mitotic spindles. It
induces the completion of mitosis and cytokinesis.
Molecular biology of the cell sixth edition (Bruce Alberts et.al)
 Proteins involved in cell-cycle control

• Cyclin-dependent protein kinases (Cdk): are the basic components of the cell cycle control
system. These proteins are periodically activated, the activity of these kinases increases or
decreases throughout the cell cycle. They lead to periodic changes in the phosphorylation of
proteins that initiate or regulate the main events of the cell cycle.
• Cyclins are proteins that are periodically synthesized and degraded in each cell cycle. These
periodic changes in cyclin proteins result in periodic activation of cyclin-Cdk complexes at specific
stages of the cell cycle. The primary determinant of Cdk activity during the cell cycle is the
increasing and decreasing cyclin levels.
• Cdk-modifying protein kinase and phosphatases: cdk-activating kinase (CAK), Wee1 kinase,
Cdc25 phosphatase
• Cdk inhibitory proteins (CKI): p27, p21, p16
• Ubiquitin ligases and their activators           
exp. APC / C: catalyzes the stimulation of regulatory proteins involved in exit from mitosis
 Cyclin-dependent kinase (Cdk) activation

-In the inactive state, without cyclin bound,

the active site is blocked by a region of the
protein called the T-loop

-The binding of cyclin causes the T-loop to

move out of the active site, resulting in
partial activation of the Cdk2.

- Phosphorylation of Cdk2 (by CAK) at a

threonine residue in the T-loop further
activates the enzyme by changing the shape
of the T-loop, improving the ability of the
enzyme to bind its protein substrates. Thus,
a fully active enzyme is formed.

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

 Cyclin-dependent kinase (Cdk) inhibition

-Cdk activity can be suppressed by inhibitory -Binding of Cdk inhibitor proteins (CKI)
phosphorilation. inactivates cyclin-Cdk complexes.

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

 Three classes of cyclins function in the cell-cycle control system

G1/S-cyclins activate G1/S Cdks in late G1 and thereby help trigger progression
through Start point, resulting in a commitment to cell-cycle entry. Their levels fall in S phase.
S-cyclins bind S-Cdks soon after progression through Start and help stimulate
chromosome duplication. S-cyclin levels remain elevated until mitosis,
and these cyclins also contribute to the control of some early mitotic
events.
M-cyclins activate M-Cdks that stimulate entry into mitosis at the G2/M transition.M-cyclin
levels fall in mid-mitosis.
Molecular biology of the cell sixth edition (Bruce Alberts et.al)
Cyclins and cyclin-dependent kinases in mammalians

Lodish
 An overview of the cell-cycle control system

-Cell cycle control system consist of a

series of cyclin-Cdk complexes.

-The activity of each complex is also

influenced by various inhibitory
mechanism.

When the mitosis phase is over, the Cdks are inactivated and entered
into a stable G1.
Molecular biology of the cell sixth edition (Bruce Alberts et.al)
Molecular biology of the cell sixth edition (Bruce Alberts et.al)
Control of S Phase

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

 Summary of the S
phase

-In the synthesis phase (S phase), both DNA molecules in the chromosomes and the chromatin
proteins are correctly duplicated.

-Chromosome duplication is triggered by activation of S-Cdks.

-S-Cdks activate proteins that will unwind the DNA double strand and initiate replication.

-Once the origin of the replication is activated, the S-Cdks inhibit proteins that will restart DNA
replication. Thus, each replication origin is activated only once in each S phase.

-As a result, S-Cdks ensure that DNA replication takes place once per cell cycle.
Mitosis

- A type of division where genetically identical 2 cells form from a parent cell.
-It is a form of reproduction in unicellular organisms. In multicellular organisms, it plays a role in
growth, development and repair of damaged body parts.

Rat fibroblasts

Rat fibroblasts undergoing

mitosis

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

Prophase

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

Prometaphase

Chromosomes

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

Metaphase

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

Anaphase

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

Telophase

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

Cytokinesis

The cell divides by the

contraction of the
contractile ring consisting of
actin and myosin filaments

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

 Control of the M phase

-Following completion of the S phase and passing the

G2 phase, the cells enter the M phase.

-The M phase starts with mitosis. The sister chromatids

in mitosis are divided into a pair of identical cells.

-After mitosis is completed, the second major event of

the M phase, cytokinesis takes place and the cell is
divided into 2 identical parts with two identical nuclei.

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

 Control of the M phase

The mitosis can be divided into 2 parts managed by

M-Cdk and APC/C components.

1. The sudden increase in M-Cdk activity at the

G2/M transition triggers the events in the early
stages of mitosis. At this stage, M-Cdk and several
other mitotic protein kinases trigger many events
related to the entry to mitosis.
2. Metaphase / anaphase transition; the APC/C
complex causes the degradation of cohesin
proteins that hold the sister chromatids together,
allowing the separation of sister chromatids and
the progression of mitosis to the anaphase.

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

M-Cdk controls many different and complex cell arrangements in early stages of mitosis

• It triggers the formation of mitotic spindle. In addition, it ensures that each sister chromatid
pair is attached to the spindle at the opposite pole.
• It triggers chromosome condensation.
• M-Cdks induce the degradation of the nuclear envelope into small vesicles by phosphorylating
the subunits of the nuclear pore complexes and the nuclear lamina in the nuclear envelope.
• They induce the rearrangement of the actin cytoskeleton and the Golgi apparatus.
• They induce the formation of spindle in prophase.
• All of these functions occur as a result of M-Cdk's activating function by phosphorylating
specific proteins, many of which have not yet been identified.
• Two protein families; Polo-like kinases and Aurora kinases also activate certain proteins in the
early stages of mitosis by phosphorylating them and control early mitotic events.
M-Cdks and other mitotic protein kinases control microtubule dynamics by
phosphorylating some regulatory proteins, including microtubule-associated
proteins (MAPs).

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

 M-Cdks phosphorylate the subunits of the condensin and cohesin
proteins and allow chromosome condensation.

Mitotic
chromosome

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

 Centriole replication - When cell enters phase S, centrioles start to duplicate.
The two centriols are separated and each centriol
creates a new centriole.

-G1 / S-Cdks (cyclin E and Cdk2 in animal cells) help the

entry to cell cycle and the start of centrosome
duplication.

-Aurora-A and Plk help the centrosome maturation by

phosphorylating the centrosome components.

-Centrosome pair remains together until the cell enters

the mitosis.

- There is a parallelity between chromosome duplication

and centrosome duplication. Both are semiconservative
and replicate once per cell cycle.
Molecular biology of the cell sixth edition (Bruce Alberts et.al)
 APC/C complex triggers the separation of sister chromatids in anaphase and
completion of mitosis

• By entering the anaphase and losing cohesins, the sister chromatids suddenly and synchronously
separate and move toward opposite poles of the mitotic spindle.

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

 The initiation of sister chromatid separation by the APC/C.

-Synthesis of Cdc20 increases while

the cell approachs mitosis.

-M-Cdk helps to bind APC/C complex

with Cdc20 by phosphorylating and
thus activation.

-Activated APC/C trigger to

degradation to securin protein.

-The separase which is activated at

the end of the metaphase by the
degradetion of securin ptotein
separates the chromatids by cleaving
the cohesins which hold the sister
Molecular biology of the cell sixth edition (Bruce Alberts et.al)
chromatids together.
 Unattached Chromosomes Block Sister-Chromatid
Separation: The Spindle Assembly Checkpoint

• checkpoint mechanism which ensures that cells do not enter

anaphase until all chromosomes are correctly bi-oriented on the
mitotic spindle is named as «spindle assembly checkpoint mechanism»
• The spindle assembly checkpoint depends on a sensor mechanism
that monitors the strength of microtubule attachment at the
kinetochore
• Any kinetochore that is not properly attached to the spindle sends out
a diffusible negative signal that blocks Cdc20–APC/C activation
throughout the cell and thus blocks the metaphase-to-anaphase
transition
 Cytokinesis
• Cytokinesis: Division of the cytoplasm starts towards the end of the telophase.

The contractile ring consists of a dynamic

combination of actin, myosin 2 filaments
and some structural proteins.

Cleavage furrow

Molecular biology of the cell sixth edition (Bruce Alberts et.al)

-Cytokinesis in the right place and at the right time
occurs by a mechanism related to mitotic spindle.

-Cytokinesis occurs in the right place because the spindle

along the anaphase generate a signal to initiate furrow
formation in a position in the middle of the spindles.

- Cytokinesis occurs at the right time because

dephosphorylation of Cdk substrates initiates cytokinesis.
 Summary of the M phase
• M-Cdks trigger events in the early stages of mitosis, such as chromosome
condensation, formation of mitotic spindle, and correct attachment of
sister chromatids to spindles.
• Anaphase is triggered by the APC / C complex. The APC / C complex triggers
the degradation of proteins that hold sister chromatids together.
Phosphorylation of the APC/C helps Cdc20 bind to the APC/C, thereby
helping to create an active complex. Among the kinases that phosphorylate
and thus activate the APC/C is M-Cdk. Thus, M-Cdk not only triggers the
early mitotic events leading up to metaphase, but it also sets the stage for
progression into anaphase.
• The APC / C complex also induces cyclin degradation and M-Cdks
inactivation.
• The Cdks become inactive as a result of the degradation of cyclins.  As a
result of the inactivation of the Cdks, the targets of the Cdks are
dephosphorylated. So, the spindles are separated, the nuclear envelope is
regenerated.
 The organelles surrounded by membrane are transferred to the
daughter cells during cytokinesis.

• The numbers of mitochondria are approximately doubled in each cell cycle and
distributed to the daughter cells.
• ER is organized by microtubules in the interphase cell. With the entry to mitosis,
the microtubules are reorganized, the nuclear envelope is degraded and the ER is
released. In most of the cells, ER is intact in mitosis and is divided into two in
cytokinesis.
• The Golgi apparatus is reorganized by disintegrating in mitosis and reformed in
telophase.