Unit 7 Cell cycle

Eukaryotic cell cycle

Mitosis and meiosis

Regulation of cell cycle

Cell cycle: the sequence of stages
through which a cell passes from one
cell division to the next

Mitosis (M): a type of cell division in

which a daughter cell receives the
same number of chromosomes as the
parent cell

Cytokinesis: the division of cytoplasm

Interphase: the time interval

between nuclear divisions when a cell
increases in mass, roughly doubles
the cytoplasmic components, and
duplicates its chromosomes

The cell cycle. The circle represents the entire life cycle of the cell, which
can be divided into two major phases: interphase and the division phase.
Most cells spend the majority of their time in interphase.
• As the cell moves through the cell cycle, chromosomes may be either
uncondensed or condensed.
• Uncondensed chromosomes are long, thin strands that cannot be seen under a
light microscope.
• A condensed chromosome can be seen under a light microscope.
• Condensed chromosomes may be either unduplicated or duplicated. In a
duplicated chromosome, the original chromosome and its duplicate are
attached to each other by a structure called the centromere.
• While attached to one another, the two chromosome duplicates are called sister
chromatids.
• Since sister chromatids contain identical genetic information, the pair, attached
at the centromere, is still considered to be one chromosome.
Fig: An unduplicated and a duplicated
chromosome
 The inner surface of the nuclear
envelope is covered with a network
of fibers that make up the nuclear
lamina. This is composed of
intermediate filament fibers called
nuclear lamins.

 Nuclear lamina gives the nucleus its

shape and is also involved in the
deconstruction and reconstruction
of the nuclear envelope that
accompanies cell division.
Interphase
• Cells spend most of their time in interphase.
• In this phase of the cell cycle, cells are not actively dividing.
• Interphase includes the G1, S, and G2 phases of the cell cycle.
• Cells in interphase grow and undergo the various metabolic processes needed for
their functioning during G1, S, and G2.
• Chromosomes are uncondensed throughout interphase.
• During G1, cells undergo a period of rapid growth, and the chromosomes are
unduplicated.
• During the S phase, cells begin to prepare for division by duplicating its
chromosomes.
• At the end of the S phase, all the chromosomes are therefore duplicated
chromosomes.
• During G2, the cell again grows and it completes the preparations for division
(mitosis, or the M phase).
M phase usually lasts only an hour or so in mammalian cells, interphase may
extend for days, weeks, or longer, depending on the cell type and the conditions.

S phase is also the period when the cell synthesizes the additional histones that
will be needed as the cell doubles the number of nucleosomes in its
chromosomes
The Stages of Mitosis

Prophase: It is the first phase of mitosis.

• The chromosomes in the nucleus become visible under a
microscope as they shorten and thicken.

Formation of the Mitotic Chromosome

• The nucleus of an interphase cell contains tremendous lengths of

chromatin fibers.
• The extended state of interphase chromatin is ideally suited for the
processes of transcription and replication but not for segregation into
two daughter cells.
• Before segregation of chromosomes, a cell converts chromatin fibers
into much shorter, thicker structures by a remarkable process of
chromosome compaction (or chromosome condensation), which
occurs during early prophase.
• Supercoiled DNA occupies a much smaller volume than relaxed DNA.
DNA supercoiling plays a key role in compacting a chromatin fiber into
the tiny volume occupied by a mitotic chromosome.

• Chromosome compaction at prophase requires topoisomerase II and

condensin which presents as part of the mitotic chromosome scaffold.

• Condensin is activated at the onset of mitosis by phosphorylation of

several of its subunits by the cyclin–Cdk responsible for driving cells
from G2 into mitosis. Thus condensin is one of the targets through
which Cdks are able to trigger cell cycle activities.

• Prior to replication, the DNA of each interphase chromosome becomes

associated at sites along its length with a multiprotein complex called
cohesin. Following replication, cohesin holds the two sister chromatids
together continuously through G2 and into mitosis when they are
ultimately separated.
Each mitotic chromosome is comprised of a pair of sister chromatids
connected to one another by the protein complex cohesin.
Colorized scanning electron micrograph of several metaphase
chromosomes showing the paired identical chromatids associated loosely
along their length and joined tightly at the centromere.
The chromatids are not split apart from one another until anaphase.
Centromeres and Kinetochores
• Primary constriction or an indentation is the most notable landmark on a mitotic
chromosome which marks the position of the centromere.
• At the centromere highly repeated DNA sequences serve as the binding sites for
specific proteins.
• Examination of sections of a mitotic chromosome reveals the presence of a
proteinaceous, button-like structure, called the kinetochore, at the outer
surface of the centromere of each chromatid.
• Most of the proteins that make up the kinetochore are assembled at the
centromere at early prophase.
• Kinetochore proteins are thought to be recruited to the centromere because of
the presence of the novel nucleosomes containing the histone variant CENP-A
at that position.
The kinetochore functions as:

(1) the site of attachment of the chromosome to the dynamic microtubules of the
mitotic spindle,

(2) the residence of several motor proteins involved in chromosome motility, and

(3) a key component in the signaling pathway of an important mitotic checkpoint.

The kinetochore. (a) Electron micrograph of a section through one of the kinetochores of a
mammalian metaphase chromosome, showing its three-layered (trilaminar) structure.
Microtubules of the mitotic spindle can be seen to terminate at the kinetochore.
(b) Schematic representation of the kinetochore, which contains an electron-dense inner
and outer plate separated by a lightly staining interzone.

The inner plate contains a variety of proteins attached to the centromeric heterochromatin
of the chromosome. Fibrous corona is associated with the outer plate which binds motor
proteins involved in chromosome movement.
(c) A schematic model showing a proposed disposition of several proteins found at the outer
surface of the kinetochore. Among the motor proteins associated with the kinetochore,
cytoplasmic dynein moves toward the minus end of a microtubule, whereas CENP-E moves
toward the plus end. These motors may also play a role in tethering the microtubule to the
kinetochore. The protein labeled “depolymerase” is a member of the kinesin superfamily that
functions in depolymerization of microtubules rather than motility. Ndc80 is a protein complex
consisting of four different protein subunits that form a 57 nm-long, rod-shaped molecule
extending outward from the body of the kinetochore. Globular domains at either end of the
complex mediate attachment to the microtubule and kinetochore. These Ndc80 fibrils have
been implicated as couplers of the kinetochore to the plus end of a dynamic microtubule
Motor proteins are molecular motors that use ATP hydrolysis to
move along cytoskeletal filaments within the cell. They fulfil
many functions within biological systems, including controlling
the sliding of filaments in muscle contraction and mediating
intracellular transport along biopolymer filament tracks.

Myosin is the motor, actin filaments are the tracks along which
myosin moves, and ATP is the fuel that powers movement.
Prophase contd.

• In animal cells as the chromosomes become visible, a small body in the

cytoplasm separates and its parts move to opposite poles of the cell.
These tiny structures, called centrioles.
• Centrioles provide attachment for the spindle fibres, which serve as
guide wires for the attachment and movement of the chromosomes
during cell division.
• Collectively, the centrioles and spindle fibres make up the spindle
apparatus.
• Most plant cells do not have centrioles, but spindle fibres still form and
serve a similar purpose.
• The centromere joining the two chromatids helps anchor the
chromosomes to the spindle fibres.
• When viewed under a microscope during prophase, the nuclear
membrane appears to fade; it starts dissolving to allow the separation of
chromosomes and cell organelles.
The stages of mitosis in an animal cell (left drawings) and a plant
cell (right photos)
Formation of the Mitotic Spindle

Cells possess two fundamentally different mechanisms—one centrosome-

dependent and the other centrosome-independent—to form mitotic
spindle.

• In animal cells, microtubule assembly is initiated by a special microtubule-

organizing structure, the centrosome. As a cell progresses past G2 and
into mitosis, the microtubules of the cytoskeleton undergo sweeping
disassembly in preparation for their reassembly as components of a
complex, micro-sized machine called the mitotic spindle.

• With the onset of S phase, as DNA replication begins in the nucleus, each
centriole of the centrosome initiates its “replication” in the cytoplasm.

• The process begins with the appearance of a small procentriole next to

each preexisting (maternal) centriole and oriented at right angles to it.
• Subsequent microtubule elongation converts each procentriole into a full-
length daughter centriole.
• During early prophase in a typical animal cell, the first stage in the
formation of the mitotic spindle is the appearance of microtubules in a
“sunburst” arrangement, or aster, around each centrosome.
• The process of aster formation is followed by separation of the
centrosomes from one another and their subsequent movement around
the nucleus toward opposite ends of the cell.
• Centrosome separation is driven by motor proteins associated with the
adjacent microtubules.
• As the centrosomes separate, the microtubules stretching between
them increase in number and elongate.
• Eventually, the two centrosomes reach points opposite one another,
thus establishing the two poles of a bipolar mitotic spindle.
• Following mitosis, one centrosome will be distributed to each daughter
cell.
The initiation of centrosome duplication at the G1–S transition is normally
triggered by phosphorylation of a centrosomal protein by Cdk2, the same
agent responsible for the onset of DNA replication.

Centrosome duplication is a tightly controlled process so that each mother

centriole produces only one daughter centriole during each cell cycle.
The formation of additional centrioles can lead to abnormal cell division
and may contribute to the development of cancer
The centrosome cycle of an animal cell

(a) During G1, the centrosome contains

a single pair of centrioles that are no
longer as tightly associated as they
were during mitosis.
(b) During S phase, daughter
procentrioles form adjacent to
maternal centrioles so that two
pairs of centrioles become visible
within the centrosome.
The daughter procentrioles continue to
elongate during G2 phase, and at the
beginning of mitosis, the centrosome
splits, with each pair of centrioles
becoming part of its own centrosome.
As they separate, the centrosomes
organize the microtubule fibers that
make up the mitotic spindle.
 Formation of a spindle pole in the absence of
centrosomes

Pole

Formation of a spindle pole in the absence of centrosomes

In this model, each motor protein has multiple heads, which are bound to
different microtubules.
The movement of these motor proteins causes the minus ends of the
microtubules to converge to form a distinct spindle pole.
This type of mechanism is thought to facilitate the formation of spindle
poles in the absence of centrosomes but may also play a role when
centrosomes are present.
Metaphase
• The second phase of mitosis is metaphase.
• Chromosomes composed of sister chromatids move toward the centre of the
cell.
• This centre area is called the equatorial plate, because, like the equator of
Earth, it is midway between the poles of the cell.
• The chromosomes appear as dark, thick filamentous structures that are
attached to the spindle fibres.
• Even though they are most visible at this stage, it is still very difficult to count
the number of chromosomes in most cells because the chromosomes are
entangled. Chromatids can become intertwined during metaphase.

Anaphase
• Anaphase is the third phase of mitosis.
• The centromeres divide and the sister chromatids, now referred to as
chromosomes, move to opposite poles of the cell.
• If mitosis proceeds correctly, the same number and type of chromosomes will
be found at each pole.
• Occasionally, segments of the chromatids will break apart, and may reattach,
in anaphase.
Telophase

• The last phase of mitosis is telophase.

• The chromosomes reach the opposite poles of the cell and begin to lengthen.
• The spindle fibres dissolve and a nuclear membrane forms around each mass of
chromatin.
• Telophase is followed by cytokinesis, the division of the cytoplasm.

Cytokinesis

• Once the chromosomes have moved to opposite poles, the cytoplasm begins to
divide. Cytokinesis appears to be quite distinct from nuclear division.
• In an animal cell, a furrow develops, pinching off the cell into two parts.
• This is the end of cell division.
• In plant cells, the separation is accomplished by a cell plate that forms between
the two chromatin masses.
• The cell plate will develop into a new cell wall, eventually sealing off the contents
of the new cells from each other.
Significance of Mitosis

• Mitosis is responsible for the development of the zygote into an adult.

• Equal distribution of chromosomes to each daughter cell.
• It is responsible for the growth and development of an individual.
• It maintains the constant number of chromosomes in all body cells of
an organism.
• Mitosis is required for asexual reproduction, vegetative propagation in
plants and also responsible for repair and regeneration of damaged
tissues.
• Mitosis helps in maintaining purity of genome as there is no
recombination or crossing over takes place.
• It is responsible for repair and regeneration of old and damaged cells
in animals e.g. gut epithelium, blood cells, etc.
An overview of the eukaryotic cell
cycle
• This diagram of the cell cycle
indicates the stages through
which a cell passes from one
division to the next.
• The cell cycle is divided into two
major phases: M phase and
interphase.
• M phase includes the successive
events of mitosis and cytokinesis.
Interphase is divided into G1, S,
and G2 phases, with S phase
shows period of DNA synthesis.
• The division of interphase into
three separate phases based on
the timing of DNA synthesis was
first proposed in 1953 by Alma
Howard and Stephen Pelc of
Hammersmith Hospital, London,
based on their experiments on
plant meristem cells.
The stages of mitosis in an animal cell (left drawings) and a plant cell (right
photos)
Meiosis
 In 1905, the term meiosis was coined from the Greek word meaning
“reduction.”

 Meiosis ensures production of a haploid phase in the life cycle, and

fertilization ensures a diploid phase.

 Without meiosis, the chromosome number would double with each

generation, and sexual reproduction would not be possible.

 To compare the events of mitosis and meiosis, we need to examine the

fate of the chromatids.
 Prior to both mitosis and meiosis, diploid G2 cells contain pairs of
homologous chromosomes, with each chromosome consisting of two
chromatids.

 During mitosis, the chromatids of each chromosome are split apart and
separate into two daughter nuclei in a single division. As a result, cells
produced by mitosis contain pairs of homologous chromosomes and are
genetically identical to their parents.
In contrast, during meiosis the four chromatids of a pair of replicated homologous
chromosomes are distributed among four daughter nuclei.

Meiosis accomplishes this feat by incorporating two sequential divisions without an

intervening round of DNA replication

• Each cell in our body, except the sex cells, contains 23 pairs of homologous
chromosomes, or 46 chromosomes in total.
• The 23rd pair of chromosomes, which determine sex in mammals, are called the
X and Y chromosomes and are only partially homologous.
• Males receive an X and a Y chromosome and females receive two X
chromosomes.

• During fertilization, a haploid (n = 23) sperm cell unites with a haploid (n = 23)
egg cell to produce a diploid (2n = 46) zygote.

• The fusion of male and female gametes restores the diploid chromosome
number in the zygote.
• The zygote will begin dividing by mitosis and will eventually become a
multicellular human baby.
The Stages of Meiosis as with mitosis, initiates with DNA replication.

The premeiotic S phase generally takes several times longer than a

premitotic S phase.

Prophase of the first meiotic division (i.e., prophase I) is typically

lengthened compared to mitotic prophase.

• For example, in the human female, oocytes initiate prophase I of meiosis

prior to birth and then enter a period of prolonged arrest.

• Oocytes resume meiosis just prior to the time they are ovulated, which
occurs every 28 days or so after an individual reaches puberty.

• Consequently, many human oocytes remain arrested in the same

approximate stage of prophase for several decades.
The first meiotic prophase is also very complex and is divided into several stages
that are similar in all sexually reproducing eukaryotes.
1. Leptotene
2. Zygotene
3. Pachytene
4. Diplotene
5. Diakinesis

The stages of prophase I

The stages of meiosis

The stages of prophase I

Leptotene, the first stage of prophase I, during which the chromosomes
become compacted and visible in the light microscope.
• Pair of identical chromatids are not completely visible.
• However, in the electron microscope the chromosomes are appeared as
composed of paired chromatids.

Zygotene, the second stage of prophase I, is marked by the visible

association of homologues with one another. This process of chromosome
pairing is called synapsis.
Electron micrographs show that chromosome synapsis is accompanied by
the formation of a complex structure called the synaptonemal complex.
• The synaptonemal complex (SC) is a ladder-like structure with transverse
protein filaments connecting the two lateral elements.

• The chromatin of each homologue is organized into loops that extend from one
of the lateral elements of the SC. The lateral elements are primarily composed
of cohesin, which binds together the sister chromatids.

• SC is not required for genetic recombination. SC primarily act as a scaffold to

allow interacting chromatids to complete their crossover activities.

Schematic diagram of the synaptonemal

complex and its associated chromosomal
fibers. The dense granules (recombination
nodules) seen in the center of the SC
contain the enzymatic machinery required
to complete genetic recombination.
Genetic recombination (crossing-over) is
occur between the DNA loops from
nonsister chromatids.
• The complex formed by a pair of synapsed homologous chromosomes is called a
bivalent or a tetrad. The former term reflects the fact that the complex contains
two homologues, whereas the latter term explains the presence of four
chromatids.

• The end of synapsis marks the end of zygotene and the beginning of the next
stage of prophase I, called pachytene, which is characterized by a fully formed
synaptonemal complex.

• During pachytene, the homologues are held closely together along their length
by the SC. The DNA of sister chromatids is extended into parallel loops.

• Under the electron microscope, a number of electron-dense bodies about 100

nm in diameter are seen within the center of the SC. These structures called as
recombination nodules as they correspond to the sites where crossing-over is
taking place.

• Recombination nodules contain the enzymatic machinery that facilitates

genetic recombination, which is completed by the end of pachytene.
The synaptonemal complex

(a) Electron micrograph of a human

pachytene bivalent showing a pair of
homologous chromosomes held in a
tightly ordered parallel array. K,
kinetochore.
(b) Schematic diagram of the synaptonemal
complex and its associated chromosomal
fibers. The dense granules (recombination
nodules) seen in the center of the SC
(indicated by the arrowhead in part a)
contain the enzymatic machinery required
to complete genetic recombination.
Genetic recombination (crossing-over) is
occur between the DNA loops from
nonsister chromatids.
• The beginning of diplotene, the next stage of meiotic prophase I, is
recognized by the dissolution of the SC, which leaves the chromosomes
attached to one another at specific points by X-shaped structures,
termed chiasmata (singular chiasma).

• Chiasmata are located at sites on the chromosomes where crossing-over

between DNA molecules from the two chromosomes had occurred.

• Chiasmata are formed by covalent junctions between a chromatid from

one homologue and a nonsister chromatid from the other homologue.

• In vertebrates, diplotene can be an extremely extended phase of

oogenesis during which the bulk of oocyte growth occurs. Thus
diplotene can be a period of intense metabolic activity.
Visible evidence of crossing-over.

• (a,b) Diplotene bivalents from the

grasshopper showing the
chiasmata formed between
chromatids of each homologous
chromosome.
• (c) Scanning electron micrograph
of a bivalent from the desert
locust with three chiasmata
(arrows).
Diakinesis is the final stage of meiotic prophase I.

• In this stage the meiotic spindle is assembled and the chromosomes are
prepared for separation.

• Diakinesis ends with the disappearance of the nucleolus, the breakdown

of the nuclear envelope, and the movement of the tetrads to the
metaphase plate.

• In vertebrate oocytes, these events are triggered by an increase in the

level of the protein kinase activity of MPF (maturation-promoting
factor).
At metaphase I, the two homologous chromosomes of each bivalent are
connected to the spindle fibers from opposite poles.
• In contrast, sister chromatids are connected to microtubules from the
same spindle pole, which is made possible by the side-by-side
arrangement of their kinetochores.

• The orientation of the maternal and paternal chromosomes of each

bivalent on the metaphase I plate is random; the maternal member of a
particular bivalent has an equal likelihood of facing either pole.

During anaphase I, homologous chromosomes separate and each pole

receives a random assortment of maternal and paternal chromosomes.

Thus, anaphase I is the cytological event that corresponds to Mendel’s

law of independent assortment.

As a result of independent assortment, organisms are capable of

generating a nearly unlimited variety of gametes.
• Separation of homologous chromosomes at anaphase I requires the
dissolution of the chiasmata that hold the bivalents together.

• The chiasmata are maintained by cohesion between sister chromatids

in regions that flank these sites of recombination.

• The chiasmata disappear at the metaphase I–anaphase I transition, as

the arms of the chromatids of each bivalent lose cohesion.

• Loss of cohesion between the arms is accomplished by proteolytic

cleavage of the cohesin molecules in those regions of the chromosome.

• In contrast, cohesion between the joined centromeres of sister

chromatids remains strong, because the cohesin situated there is
protected from proteolytic attack.

• As a result, sister chromatids remain firmly attached to one another as

they move together toward a spindle pole during anaphase I.
(a) Schematic diagram of a pair of homologous chromosomes at metaphase
I. The chromatids are held together along both their arms and
centromeres by cohesin. The pair of homologues are maintained as a
bivalent by the chiasmata.
(b) At anaphase I, the cohesin holding the arms of the chromatids is
cleaved, allowing the homologues to separate from one another.
Cohesin remains at the centromere, holding the chromatids together.
Telophase I of meiosis I produces less dramatic changes than telophase of
mitosis.

• Although chromosomes often undergo some dispersion, they do not

reach the extremely extended state of the interphase nucleus.

• The nuclear envelope may or may not reform during telophase I.

Interkinesis

The stage between the two meiotic divisions is called interkinesis and is
generally short-lived.

In animals, cells in this fleeting stage are referred to as secondary

spermatocytes or secondary oocytes.

These cells are characterized as being haploid because they contain only
one member of each pair of homologous chromosomes.
• Even though they are haploid, they have twice as much DNA as a
haploid gamete because each chromosome is still represented by a
pair of attached chromatids.

• Secondary spermatocytes are said to have a 2C amount of DNA, half as

much as a primary spermatocyte, which has a 4C DNA content, and
twice as much as a sperm cell, which has a 1C DNA content.

Interkinesis is followed by prophase II.

• During prophase II, nuclear envelope broken down again, if it

had reformed in telophase I.

• The chromosomes become recompacted and line up at the

metaphase plate.
Metaphase II
Unlike metaphase I, the kinetochores of sister chromatids of metaphase II
face opposite poles and become attached to opposing sets of
chromosomal spindle fibers.

The progression of meiosis in vertebrate oocytes stops at metaphase II.

• The arrest of meiosis at metaphase II is brought about by factors that

inhibit APCCdc20 activation, thereby preventing cyclin B degradation.

• As long as cyclin B levels remain high within the oocyte, Cdk activity is
maintained, and the cells cannot progress to the next meiotic stage.
Metaphase II arrest is released only when the oocyte (now called an
egg) is fertilized.

• Fertilization leads to a rapid influx of Ca2+ ions, the activation of

APCCdc20, and the destruction of cyclin B.
• The fertilized egg responds to these changes by completing the second
meiotic division.
Anaphase II begins with the synchronous splitting of the centromeres,
which had held the sister chromatids together, allowing them to move
toward opposite poles of the cell.

Telophase II
Meiosis II ends with telophase II, in which the chromosomes are once again
enclosed by a nuclear envelope.

The products of meiosis are haploid cells with a 1C amount of nuclear

DNA.
Separation of homologous chromosomes during
meiosis I and separation of chromatids during
meiosis II.
(a) Schematic diagram of a pair of homologous
chromosomes at metaphase I. The chromatids
are held together along both their arms and
centromeres by cohesin. The pair of
homologues are maintained as a bivalent by
the chiasmata.
(b) At anaphase I, the cohesin holding the arms of
the chromatids is cleaved, allowing the
homologues to separate from one another.
Cohesin remains at the centromere, holding
the chromatids together.
(c) At metaphase II, the chromatids are held
together at the centromere, with microtubules
from opposite poles attached to the two
kinetochores. The kinetochores of the sister
chromatids are now on opposite sides of the
chromosome, facing opposite poles.
(d) At anaphase II, the cohesin holding the
chromatids together has been cleaved,
allowing the chromosomes to move to
opposite poles.
Cell cycle control system
Cell cycle control system directed the sequential events of the cell cycle.

Cell cycle is regulated by checkpoints by both internal and external controls.

A checkpoint in the cell cycle is a critical control point which decides weather
a cell stop and go ahead in a cell cycle. It involves modulation of stop and go-
ahead signals.

There are three major check-points in the cell cycle, G1, G2, and M-phase.

If a cell receives a go-ahead signal at G1 check point, it will usually completes

the S, G2 and M phase and divide.

Or, if it does not receive a go-ahead signal at that point, cell will exist the
cycle and entered into a non-dividing state called the G0 phase.

Most cells of the human-body are actually in the G0 phase ex. Mature nerve-
cells and muscle cells never divide.
Molecular basis for the cell cycle control

This include regulatory proteins, which are of two types:

1. Kinases
2. Cyclins

Protein kinases are enzymes that activate or inactivate other proteins by

phosphorylating them.
Particular protein kinases give the go-ahead signals at the G1 and G2
checkpoints.

In a growing cell, kinases that involved in the cell cycle are present in a
constant concentration but in a inactive state.

For their activation, they attached with cyclin.

Cyclin is a protein and gets its name from its cyclically fluctuating
concentration in the cell.
As these kinases requires cyclin for their activation they are called as cyclin-
dependent kinases or cdks.

The activity of cdk rises and falls with the change in concentration of cyclin.

Cyclin-cdk complex is known as Maturation promoting factor (MPF).

So, MPF consists of two subunits

1. Have kinase activity transfers phosphate group from ATP to specific serine
and threonine residues of specific protein substrates and
2. a regulatory subunit called cyclin

When the cyclin concentration is low, the kinase lacks the cyclin subunit and,
as a result, is inactive.
When the cyclin concentration rises, the kinase is activated, causing the cell
to enter M phase.
These results suggested that
(1) progression of cells into mitosis depends on an enzyme whose sole
activity is to phosphorylate other proteins, and
(2) the activity of this enzyme is controlled by a subunit whose
concentration varies from one stage of the cell cycle to another

Fluctuation of cyclin and MPF levels during the cell

cycle
MPF performs several functions to control cell cycle.

1. MPF acts both directly as a kinases and indirectly by activating other

kinases.

Ex. MPF causes phosphorylation of other proteins of nuclear lamina-which

promotes fragmentation of the nuclear envelop during prometaphase of
mitosis.

2. MPF also involved in the condensation of chromosomes and spindle

formation during prophase.
To study genetic control of cell cycle two species of yeast cells were used

Budding yeast, Fission yeast

Saccharomyces cerevisiae Schizosaccharomyces pombe

reproduces through the reproduces by elongating itself and then

formation of buds at one end of splitting into two equalsized cells
the cell.
Cdk found was cdc28 Cdk found was cdc2
At both the checkpoints G1 and G2 cdk
used is cdc 2 but involves attachment
with different cyclins.
Fission yeast contd.

First transition point called START, occurs in late G1 phase involves

activation of cdc2 by one or more G1/S cyclins, once START point clear,
cell will ultimately completes cell cycle.

Second checkpoint-Passage from G2 to mitosis requires activation of cdc2

by a different group of cyclins—the mitotic cyclins

Third checkpoint-middle of M-phase, which determines whether cell will

complete cell division and reenter G1 of the next cycle.

Exit from mitosis and entry into G1 depends on a rapid decrease in Cdk
activity that results from a plunge in concentration of the mitotic cyclins
Model of cell cycle regulation in fission yeast

START G2-M transition

1 checkpoint 2 checkpoint 3 checkpoint-
sharp decline in
mitotic cyclins
Checkpoints are surveillance mechanisms that halt the progress of the cell
cycle if

(1) any of the chromosomal DNA is damaged, or

(2) certain critical processes, such as DNA replication during S phase or
chromosome alignment during M phase, have not been properly
completed.

Checkpoints ensure that each of the various events that make up the cell
cycle occurs accurately and in the proper order.
Damage to DNA also leads to the synthesis of proteins that directly inhibit the
cyclin–Cdk complex that drives the cell cycle.

For example, cells exposed to ionizing radiation in G1 synthesize a protein called

p21 (molecular mass of 21 kDa) that inhibits the kinase activity of the G1 Cdk.

This prevents the cells from phosphorylating key substrates and from entering S
phase.

p53 involvement in control of cell growth

p53 is normally very short-lived, but phosphorylation by Chk2 (other checkpoint

kinase called Chk2) stabilizes the protein, enhancing its ability to activate p21
transcription. Once transcribed and translated, p21 directly inhibits the Cdk.

Cdk Inhibitors Cdk activity can be blocked by a variety of inhibitors. In budding yeast, for
example, a protein called Sic1 acts as a Cdk inhibitor during G1.

The degradation of Sic1 allows the cyclin–Cdk that is present in the cell to initiate DNA
replication.
Functions of Meiosis

1. Production of haploid (n) gametes: so, that fertilization restores the

normal somatic (2n) chromosome number.
2. Production of tremendous amounts of genetic variation.
3. Segregation of the two alleles of each gene. This take place due to
pairing between the two homologues of each chromosome and their
separation at the first anaphase.
4. Recombination between linked genes due to crossing over during
pachytene stage.
5. Meiosis facilitates segregation and independent assortment of
chromosomes and genes.
6. In sexually reproducing species, meiosis is essential for the continuity
of generation. Because meiosis results in the formation of male and
female gametes and union of such gametes leads to the development
of zygotes and thereby new individual.