UNIT 16 CELL CYCLE AND ~ 0 ~ 1 s

Structure
16.1 Introduction
Objectives
16.2 Cell Cycle
Interphase-The GI, S and G2 Phases
G 1-The most Variable Period of the Cell Cycle
Events Occur at Defined Stages of Ccll Cycle
Determination of Cell Cycle Times
16.3 Regulation of Cell Division
Control of the Cell Cycle in Dividing Cells
Factors that Stimulate Cell Division
16.4 Mitosis
Chromosomal Events
Role of Spindle
Chromosomal Movements
16.5 Summary
16.6 Terminal Questions
16.7 Answers

16.1 INTRODUCTION
In the previous block you have studied about the structure of nucleus and the various
processes occurring within it. Nucleus contains chromosomes which contain DNA
and proteins. You have studied how DNA and proteins form complexes and how
DNA ultimately'synthesises protein by an intermediate RNA. Also, you have read
about replication and the flow of informations from nucleus to cytoplasm, for the
genetic continuity in Block 3.
In higher eucaryotes life starts with the formation of a fertilized egg cell of
approximately 100 urn in diameter leading to the formation of trillions of cells with a
greater mass. As more and more cells are added as a result of successive divisions the
developing embryos undergo a process of differentiation and adult structures are
organised. During this period, the rate of cell division and the number of dividing
cells change so that there are fewer cells dividing in a fully formed organism than
during the course of development. In an aduli organism there are tissues where no
cells are dividing and are mitotically inactive, while there are other tissues wherein
cells die and are replaced by new cells. There are also some tissues like bone marrow
or germinal tissues or meristematic regions in plants where continuous cell division
takes place. The cells divide by two specific methods known as 'mitosis' and
'meiosis'. In this section we shall describe the events associated with the process of
cell division known as mitosis. You will read about meiosis in the next unit.

Objectives
After reading this unit you should be able to:
a recall the rate of observed cell division in some cell types and relate it to
generation/celli'cycle time
a define and use in correct context a i d terms 'cell cycle9/'generation' time, mitotic
index, S, G I , GO, 6 2
a state the four factors that are considered important in controlling the cell division
outline methods to determine different phases in a cell cycle
a explain in brief as to how the cell cycle control is studied
summarise the logic for the minimum cell cycle time
a distinguish the different phases of mitosis and draw diagrams of the same
a list the functions of spindle
give causes for chromosomal mollements during mitosis.

16.2 CELL CYCLE,

A growing cell undergoes a cell cycle that comprises essentially two periods:
(1) interphase i.e. the period of non-apparent division and (2) the period of division
Cell Division, Cell
Movement and
Differentiated Cell Types
% nerve cells and you will be amazed to know that the mammal nerve cells do not
(Fig. 16.1). In eucaryotes, division generally takes place by mitosis or meiosis. For
many years, cytologists were concerned mainly with the period of division in which
dramatic changes visible under the light microscope could be observed, and
interphase was considered a "resting" phase. Cells, however, spend most of their life
span in interphase. The interphase is a period of intense biosynthetic activity in whick
the cell doubles in size and duplicates precisely its chromosome complement. Some
differentiated cell types divide rarely such as red blood cells, skeletal muscle cells ant
divide at all after birth. Thus, for a human neuron the interphase period lasts the
entire life time of a person.
Mitosis

Fig. 16.1 The cell cycle. Time intervals are relative; actual time
varies with the cell type and species
Therefore, the cell cycle represents a complex series of stage by which cellular
material is divided equally between daughter cells. Cell division is only the final
phase. Before, the cell divides by mitosis, its main molecular components have
already been duplicated. In this respect, cell division can be considered as the final
separation of the already duplicated molecular units.

16.2.1 Interphase -The GI, S and 6 2 Phases

The life of the cell is mostly spent in interphase, a stage between the two successive
cell divisions. The introduction of cytochemical methods such as the feulgen stain,
followed by cytophotometric quantitative assay and radioautography, for the first
time, suggested that the doubling of DNA takes place during interphase. These
studies demonstrate that the synthesis occurs only in a restricted portion of the
interphase stage, the so called S-period or synthetic period which is preceded and
followed by two "gap" periods in G 1 and G2. There is no DNA synthesis between
G 1 and G2 phase of cell cycle which is divided into four successive stages: S is the
period of DNA synthesis and G2, the interval between the end of DNA synthesis
and the start of mitosis.

During G2 a cell contains twice the amount of DNA present in the original diploid
cell. Following mitosis the daughter cells again enter the G1 period and have a
equivalent DNA content.

16.2.2 GI-The Most variable Period of the Cell Cycle

GI. is the time interval between mitosis and the beginning of S-phase, which is
associated with the growth and increased activity of enzymes specific for DNA .
synthesis. The duration of the cell cycle varies greatly from one cell to another. For
mammalian cell growing in culture with a generation time of 16 hours, the different
periods would be as follows: G1=5 hours, S-7 hours, G2-3 hours and mitosis=l
hour. Generally, the S, G2 and mitotic periods are relatively constant ip the cells'of
same organism. The G 1 penod is the most variable in cell life. Depending on the Cell Cyde and Mitosis
physiological condition of the cells it may last days, months or years.

The regulation of thcdmauon of the cell cycle occurs primarily b:. arresting it at a
specific point of G 1 and the cell in the arrested condition is said to be in the GO
state. In the GO state the cell may be considered to be withdrawn from the cell cycle
(Fig. 15;l). When conditions change and growth is resumed the cell reenters the G1
pea.
Eucaryotic chromosomes undergo condensation-decondensationprocess during
interphase (Fig. 16.2).

Fig. 16.2 The condensation-decondesation cycle of chromosomes. G1 chromosomes are completely

dispe~ed; S, duplication occurs; and G2 condensation starts. A t metaphase, M and anaphase, A, the
condensation is maximal and the two centromeres the clearly visible.

The DNA of procaryotes is not processed in this way and this forms a major
difference between eucaryotes and procaryotes. The DNA of procaryotes is
replicated continuously. Most protein and RNA synthesis occurs through the
interphase.

16.2.3 Events Occur at Defined Stages of the Cell Cycle

If the biochemical events that occur at defined stages of the cell cycle (Fig. 16.3), the
most noticeable one is DNA synthesis. S-phase cells contain a factor that induces
DNA synthesis. This has been demonstrated by cell fusion experiments, in which one
set of DNA replication in G 1 nuclei can be accelerated by fusion with S-phase cells.
It is important to note here that G2 nuclei do not respond to this factor, indicating
that some mechanism must exist to block reinitiation of DNA synthesis within the
same cell cycle. Evidences for a factor that includes DNA synthesis comes from
multi-nucleated cells, such as the plasmodium of the slime mold P h p m m
polycephalum, in which all the nuclei start DNA synthesis simultaneously.

S-phase lasts for several hours, during which many units of replication are
sequentially activated. In all cells, the more condensed, heterochromatic regions of
the chromosomes replicate late during S-phase. The centromeric heterocliromatin
which contains satellite DNAs, replicates later than the rest of the chromosome.
Similarly, the X-chromosome that becomes inactivated in mammalian females is
heterochromatic and late replicating, whereas the one that remains activated is
euchromatic and replicates earlier, about which you have read in Unit 13. Molecular
events linked to the cell cycle, including the synthesis of histones during S-phase, .the
decrease in protein synthesis that occurs during mitosis; the decrease in C-AMP
levels during mitosis, phosphorylation of histones (especially HI) during chromatin
condensation and many other events.
The most important point in the regulation of the cell cycle occurs in the G 1 phase,
during which it must decide whether the cell will start a new cell cycle or becomes
arrested in the GO state. Once this G 1 checkpoint has been passed, the cell goes on
to complete a new cycle.
Cell Division, Cell f f h s t of RNA synthesis and
Movement and
Differentiated Cell Typeb

Fig. 16.3 Molecular events during the cell cycle

16.2.4 Determination of Cell Cycle Times

. How does one deterdine the length of each phase of the cell cycle? Fist it is
necessary to establish the length of the total cycle, which can easily be done in a
homogeneous population of cultured cells by periodically counting the number&
cells present under a microscope and recording the number of hours required for t1:e
total cell number to double. Alternativelv. the total cell mass can be monitored. 0 nce
this interval is known the length of the ~Iphasecan be estimated by adding titrated
thymidine to the tissue culture medium for a brief period that is much shorter than
the S-phase itself (typically 30 minutes or less). The cells are then preserved for
autoradiography and the fraction of the cells that have incorporated the radioiso:t+pc
into their b ~ ~determined
i s by counting the fraction of cell with exposed silver -
grains over their nucleus. As explained in Figure 16.4, this fraction multiplied by the
total cell cycle time is roughly equal to the average length of the S-phase in the
population.

I
-24 hours- -X24=2 how
I2
determination of the length
of M phase by the mitotic M phase = 2 hours X c o d o n factor
index
000@0090@@0@ab

-
-I
determination of the kngth
of S phase by autoradiography
-24

of ('H)thymidive - labeled ceb O 8 D 8 0

t
Lours
S

0
Q Q Oab

>-sruoh
48
1
-X24=8hours
4
I2
S phase = 8 hours = w d o n factor

@ 18
-X48=8hours
4
U
s phase = &hoursx wrrsction factor

~ig.-16.4Illustration of the general principle that the length of each phase of the cell cycle is
approximately equal to the fraction of the cells in that phase at any instant multiplied by the total cell
cycle time. This calculation is based on the assumption that all the cells in the population are growing at
the same rate. The indicated 'correction factors' range from 0.7 for early G1 cells to 1.4 for mitoticcells
with an intermediate value for S-phase cells. A correction factor is needed because there are always more
young cells than old cells in a continuously dividingpopulation. The exact age distribution '9 given by the
equation y=2-I where y varying from 2 to 1 is the relative number of cells at cell cycle age x (where x
varies from 0 for the earliest G I cells to 1 for the latest M-phase cells).
 The length of M-phase can be determined in an analogous fashion by scanning the Cell Cycle end Mitosis
II cell population by light microscopy and determining the fraction of cells containing
condensed chromosomes at any one time since the last cell division. For example, a
single old cell becomes two young cells as soon as it divides, and there will be twice
as many cells in early G 1 as in late M-phase.

Cell cycle analysis has been made much easier by the use of a fluorescene-activated
I
cell analyser. With the help of this analyser one can rapidly determine the relative
fluorescene of a large number of cells, and their relative amounts of DNA.Those
cells with the least amount of DNA are in G I phase, those with double this amount
are either in G 2 or M, while cells in S have intermediate amounts (shaded area in
Figure 16.5).

I cells in GI
phase.
I
I

Cells in GZand.
M phases
I

0 1 2
relative amount of DNA per cell
(arbitary units) '
Fig. 16.5 Typical results obtained for a growing cell population when the DNA content of its individual
cells is determined with a fluorescene analyser. DNA amounts are given om arbitrary units and are
determined by the amount of fluorescent dye bound. The fact that the number of cells in G 1 is much
greater than the number of cells in G2 and M indicates that the G1 phase is longer than G2 phase in this
population.

The length of G I , and G 2 plus M and S-phase of the cell cycle can be readily
calculated (Fig. 16.4).
Cell Mvimion, Cell
Movement -8
M f l e m t b t c d Cell T+

16.3 REGULATION OF CELL DIVISION

You have studied till now the cell cycle and cell division. The manner in which
mitosis is regulated is yet to be understood. However, cell division regulation
encompasses two fundamental questions: firstly, by what mechanism the cell cycle in
dividing stage is controlled in cells, and secondly by which mechanisms external
agents trigger the onset of cell division in cells that are not actively dividing.

16.3.1 Control of the Cell Cycle in Dividing Cells

As pointed out in the earlier section cells generally shorten or lengthen the times
between successive cell divisions by shortening or lengthening the time spent in G1
stage. The regulation of the eucaryotic cell cycle is intimately associated with events
during the G 1 phase. Direct evidence for the ithportance of G 1in regulating the cell
cycle has come from studies that limit cell growth, such as lack of nutrients or
appropriate growth factors. Under such conditions, cells stop growing at sl?ecific
point in the cell cyclk situated near the end of G1 called the restriction point, and
'
then enter into a special phase called GO, where they may exist for long periods
outside the normal cycle of cell division.
The cell fusion experiments reveal that cytoplasmic factors present in S-phase cells
are capable of triggering the premature entrance of G1 nuclei into S-phase. Such
observations suggest that the production of specific substances during 91 is
responsible for triggering the entry into S - p h i .
Yeast cells have helped us in understanding molecular event that trigger the
transition from G1 to S-phase. Yeasts we powerful tool for investigating the cell
cycle because of the ease of production of genetic mutants which a n be
characterised. Such studies have shown that a small group of genes controlling arl
event occur in GI, called start, this represents the point when ~ l l &come
s
committed to entering the mitotic cell cycle. Cells eater into a programme leading tc
S-phase, mitosis and cell division. The genes found to be essential for the start
junction, codes for a protein resembling a protein kinase (an enzyme catalysing the
phosphorylation of other proteins). .
In bacterial cells commitment to the cell diVision'cycle requires the production of Cell Cycle and Mitosis
DNA binding proteins that dest.abilise the double helix and allowing the enzymes
involved in replication to have direct access to the DNA. Proteins capable of
destabising the DNA double helix are involved in the initiation of DNA replication
in eucaryotic cells about 'which you have studied in Block 3. ?he meclianism
responsible for unpacking this nucleosomal organisation is not known in detail, but it
is thought that phosphorylation of histone molecules by protein kinase occurs during
S-phase. Histone phosphorylation might be an essential requirement for the entrance
of the cell into S-phase.

Although G1 phase appears to be the main site of regulation of the cell cycle in
euuryotes, some cells stop their cyclein G2. Such G2-arrested cells arecapable of
quicklyfitiating mitosis in response to an appropriate stimulus, and therefore occur
in places such as skin where rapid cell division may be required in response to an
injury. In contrast to G2-arrested cells, cells in either GO or G1 require many hours
to enter mitosis because of the necessity of first passing through the remainder of GI,
S, and G2. The completion of the cycle from G1 to G2 varies from 8-20 hours in
actively growing animal and plant cells.
.
16.3.2 Factors that Stimulate Cell Division
In addition to the importance of metabolic events occurring within the cell, external
conditions also exert controls over the process of cell division. In procaryotic cells the
major controlling factor is the presence of sufficient nutrients in the external
environment. As long as adequate nutrients are available, cell division generally
continues. In eucaryotic cells, which are typically multicellular organkps, division of
each cell must be carefully regulated to suit the needs of the organism as a whole. A
variety of external factors are involved in coordinating the division of such cells.
Certain steroid hormones such as testosterone and estrogens have b&n found to
stimulate the.growth and division of specific types of animal cells; while hormones
like auxins, cytokjnins, and gibberellins stimulate the divis'ion of plant cells. Although
the preceding types of hormones act by entering directly into thew target cells, other
growth promoting factors have been identified which bind to specific receptors in the
plasma membrane. Well known examples of such growth factors include epidermal
wwtb factor, platelet-derived growth factor, nerve growth factor, thymosin and
ewhropoietin. All these proteins stimulate the proliferation of a s p d c group of
target cells.
Although the mechanism by which such molecules stimulate cell division is not
completely,understood, progress hai been made in unravelling some of the early
steps invohed. One of the more thoroughly investigated examples involves the
response of cultured cells to the addition of epidermal growth factor (EGF), a
polypeptide of approximately 6400 molecular weight. Experiments mth radioactive
EGF have revealed that this pol tide 6rst binds to specific membrane receptors
located on the surface of the cell'grp
eng stimulated & divide. These receptors which
'ae initially distributed randomly over the cell surface, become clustered into patches
after binding to EGF and are subsequently taken into the cell by endocytosis.

One of the 6rst biochhical changes observed in cells treated with EGF is an
increased rate of phosphorylation of tyrosine residues located in certain intracellular
proteins as well as the EGF receptor itself. The enzymatic activity responsible for
catalysing this tyroshe-specific phosphorylation reaction resides in a portion of the
EGF receptor that hce* interior. In other words the EGF receptor, a
trans-membrane pr~tein,hinds to EGF molecules at the outer surface of the plasma
membrane and it ~ c t i o n as
s a tyrosine-specific protein kinase at the inner surface of
the plasma membrane (Fig. 16.6). Binding of EGF to the membrane receptor causes .
.
its protein kinase to become activated. The ability of certain tumor viruses to
transform normal cells into cancer cells also involves the production of a tyrosine
specific protein kinase.

The above informatioh indicates that the interaction of external factors with the
plasma membrane can trigger intracellular changes by altering the activity of an
enzyme localised within the plasma membrane. The binding of any protein hormones
to the plasma membrane triggers activation of the membrane-bound enzyme
adenylate cyclase which catalyses the formation of 3,s-cyclic adenosine
monophphate (cyclic AMP). IntraceWar concentration of cyclic AMP has been

2
found to change during e normal cell cycle as well as in response to agents that
stimulate or inhibit cell vision. Jnl;-.:ellular calcium can also trigger the onset of cell
division with the help of protein calmodulin.
c 4 DhYDI, cel
Mov- .nd
~emumtedCell Types

} H u m . membrane
I
Cell exterior
I
Fig. 16.6 Summary of the mechanism by which epidermal growth factor (EGF) influences protein
phosphorylation. Binding of EGF influences protein phosphorylation. Binding of EGF to the exterior
surface of its plasma membrane receptor cause a conformational change in the receptor that permits it to
function as a tyrosine-specific protein kinase at the cytoplasmic surface of the plasma membrane. This
enzymatic activity in turn catalyses the phosphorylation of the amino acid tyrosine (tyr) in specific
cytoplasmic proteins.
 Cdl Cycle a d Mitosis
16.4 MITOSIS
Mitosis is a complex process which splits the chromosomes equally into two daughter
cells. It involves maintenance of chromosomal continuity and diploid number. The
continuity of the chromosomal set is maintained by a special type of cell division
which is called mitosis. At the time of cell division the nucleus becomes completely
reorganised. In a somatie cell the nucleus divides by mitosis in such a way that each
of two daughter cells receives exactly the same number and kind of chromosomes
that the parent cell had.

66.4.6 Chromosomal Events

Figure 16.7 represents two pairs of homologous chromosome in a diploid nucleus.
Each chromosome duplicates itself sometime during interphase before the visible
mitotic process begins. Although mitosis accounts only for a short period of time
relative to the total time between cell division, the remaining portion of the cell cycle
is termed interphase. The nucleolus is the only structure that is visible under the light
microscope.

I) Interphase
During interphase, chromosomes are present in a different network of chromatin
that is not visible under the light microscope as an individual i.e. DNA-protein
complexes called chromatin are dispersed throughout the nucleoplasm. The events
during mitosis that follow unfolding are conventionally divided into four sub-stages:
prophase, metaphase, anaphase and telophase (Figure 16.7).
(a) Interphase
(b) prophase (c) Middle prophase (d) Late prophase
Centriole

(e) Metaphase ( f ) Early anaphase (g) Late anaphasc

(h) Telophase (i) Interphase

Fig 16.7.The stages of mitosis and cytokinesis in an animal cell

Prophase marks the beginning of the cell cycle in which condensation of chromatin
fibres occurs in discrete and visibly separate chromosomes. During prophase, each
chromosome consists of two identical units termed as sister chromatids each of
which contains identical daughter DNA molecules that were produced in the
S-phase. As firophase advances the chromatids become more condensed, owing to
the packing of the nucleoprotein fibres. Sister chromatids are held together at their
centromeres or constricted regions about which you have read in 10+2 course. At
this time, the small cylindrical centrioles begin to play a key role. As the centrioles
move apart, they generate fibrous microtubules which radiate from them in all
Cell Division, Cell directions. Centrioles, which are themselves formed of microtubules, duplicate during
Movement and interphase by forming daughter centrioles at right angles to their own positions. Some
Differentiated Cell Types of these microtubules connect the centrioles with the kinetochores, granular regions
that are attached to the centromeres of the chromatids. Other microtubules form a
network that links the centrioles as they move apart. These microtubules, together
with associated fibres and proteins, are called the'spindle. The regions surrounding
the centrioles, from which the microtubules radiate, are termed the poles or polar
regions of the cell. At the end of prophase the nuclear membrane and nucleolus
disappear, and each chromatid remains attached to spindle microtubule at its
centromere.

11) Early Prophase

The centrioles begin m o ~ n gtowards opposite poles of the cell; the chromosomes can
be seen as long threads, and the nucleus-is dispersing and becoming less distinct.

III) Middle Prophase

Chromosomes condensation is completed; each chromosome is composed of two
chromatids held together at their centromeres. Each chromatid contains one of the
two newly replicated daughter DNA molecules. The microtubular spindle begins to
radiate from the regions just adjacent to the centrioles, which are moving close to .
their poles.

IV)Late Prophase -
The centrioles reach the poles, and some spindle fibres extend from pole to the
centre or equator of the cell. Other spindle fibres extend from the poles to the
chromatids and attach to the kinetochores, which are near the centromeres of the
chromosomes. The nuclear membrane begins to disperse and disappear, and the
nucleolus is not visible.

During metaphase, the condensed sister chromatids which are connected to each pole
of the cell by the microtubules attached to their kinetochores, and migrate to the
equatorial plane of the cell. The microtubules attached to the chromosomes appear to
play a role in orienting them. It is the time when chromosomes appear thick and
discrete and can easily be identified, and counted and photographed for studies of
chromosomal abnormalities.

V) Metaphase
The chromosomes move towards the equator of the cell, where they become aligned
in the equatorial plane. The sister chromatids have not yet separated.

VI) Early Anaphase

The two daughter chromatids separate. Each contains a centromere that is linked by
a spindle fibre to one pole. Led by the centromere, each chromosome begins to move
towards the pole'to which it is attached. Simultaneously, the cell elongates as do the
pole-to-pole spindles.

VII) Late ~ n a p h a s e
Each set of chromosomes (the daughter chromatids) moves closer to its pole and
cytokinesis begins as the cleavage furrow starts in animal cells (Fig. 16.8).

b'imaEJ Cleavage furrow

I Plant cell Phragmoplast Cell plate I

Fig. 16.8 Diagram summarizing the difference between the mechanism of cytokinesis in animal and plant "
cells. In animal cells invagination of the plasma membrane proceeds from the periphery of cell towards
the interior; in plant cells the phragmoplast begins to form in the centre of the cell and then expands
towards the periphery.
Anaphase is marked by the separation of the two sister chromatids at their Cell Cy+ a d Mitoeis
centromeres. This happens simultaneously to the entire set of chromosomes. The .
members of each chromatid pair now exist as independent chromosomes, that
migrate to opposite poles of the.cel1, with each daughter cell'having t$e same
chromosome complement. The movement towards the two poles is due to a
shortening of the microtubules attached to the kinetocliores.

VIII) Telophase
It is the firlal stage of mitosis and reaches in interphase stage. New nuclear membrane
is formed around each daughter nucleus and when the chromosomes uncoil ind
become less distinct, the nucleolus becomes visible again. Throughout mitosis the
"daughter7'centriole at each pole has continued to grow until it is fully formed. At
telophase the duplicate of each of the original centrioles is completed and each of the
two centrioles at each pole begins to generate a new daughter centriole at right angles
to it.

When cflokinesis is nearly complete, and the spindle disappears as the microtubules
and other fibres depolymerise, the nuclkmli reappear.

16.4.2 Role of Spindle

In most eucaryotic cells, chromosome movement is dependent on the presence of the
mitotic spindle. This spindle apparatus contains atleast three different types of
microtubules (1) astral microtubules, which radiate from the centrioles that are only
found in animal cells, (2) kinetochore microtubules, which attach to the
chromosomal kinetochores, and (3) inter polar microtubules, which span the area
between the two spindle poles and~accountfor the bulk of spindle's structure
(Fig. 16.9).
Interpolar rnicrotubules
one microtubule

mi,crotubule centriole

Fig. 16.9 Diagrammatic model of the spindle apparatus

illustrating the three types of spindle microtubules
The assembly of spindle is regulated by changes in the equilibrium between the
polymerised and unpolymerised states of tubulin. Jt was found that exposing cells to
low temperature, high pressure, or the drug colchicine causes breakdown of the
mitotic spindle and the cessation of mitosis. However, the spindle spontaneously
reforms, as soon as the temperature is raised, the pressure lowered or colchicine
removed. This rapid reappearance of the spindle occurs even when inhibitors of
protein synthesis are present, indicating that the assembly of spindle is not dependent
on the synthesis of new spindle proteins. Calcium ions play an important role in the
process of microtubule assembly/disassembly during normal mitosis. Micro injection
of ca++into metaphase cells has been found to hasten the onset of anaphase while
conditions that lower the cytoplasmic ca++concentration show the progression into
anaphase. The mitotic spindle has been found to contain large amounts of
calmodulin, a calcium binding protein that mediates the regulatory effects of calcium
ions upon a variety of cellular processes.
Centrioles usually replicate during the S-phase of the cell cycle yielding two pairs of
centrioles that migrate to opposite sides of (he nuclear envelope at the time of
prophase. Careful replication is not a true division but rather a sequence of events in
which each pre-existing centriole induces the formation of a new centriole.
At the beginning of prophase, the two pairs of centrioles and their associated asters
begin to separate and move towards opposite sides of the cell. At the same time,
svindle microtubules start forming between the centriole pairs. By the time centrioles
Cell Divbhl, Ceu 'have reached opposite sides of the cells, the spindle microtubules are almost
MovcwM .ad complet~lyassembled. Microtubules exhibit an intrinsic polarity, one end functioning
Mfferemtbted Cell Trpes
in tubulin assembly and the other functioning in disassembly. Centrioles themselves
are not directly involved in generating the mitotic spindle.

16.1.3 ~hromosomalMovements
Several theories have been prodosed to explain the mechanism of the movement of
the two sets of chromosomes to opposite poles of the spindle during anaphase. Two
distinct types of movements occur at this time: migration of. the chromosomes
towards the poles and movement of the two poles away from one another. Several
observations suggest that the spindle microtubules play a central role in these two
types of movements. The kinetochore microtubules are the only external structures
that appear to be attached to the moving chromosomes. The kinetochore region of
the chromosome leads the way during chromosome movement, suggesting that the
chromosome is being pulled towards the pole by the microtubu!es attached to the
kinetochore. The chromosome movements can be inhibited by exposing cells to
microtubule disrupting agents such as colchicine.
-
A general description of three types of interactions for chromosome movements is
presented below:
i) Miaotubule-Microtubule Interactions
You also know that ciliary and flagellar movements are based on the active sliding
movement that is driven by the transient formation and breakage of bonds between
adjacent microtubules. The reaction involves the ATP hydrolysing protein, dynein.
The chromosome separation during mitosis is driven by the two sets of forces: a
dynein-based, ATP-requiring process of microtubule sliding that pushes the spindle
poles apart, and a separate ATP independent mechanism for pulling the
chromosomes towards the pole.
ii) Microtubule Polymerisation/Depolymerisation
Although the microtubule sliding plays a role in chromosome movement, other
factors also play a role. First the overall length of the interpolar microtubule increasef
as the two poles of the spindle separate from one another, and secondly the
individual kinetochore microtubules shorten as the chromosomes move towards the
pole. These changes suggest that interpolar microtubules are being lengthened by the
addition of tubulin subunits to their polyrnerising ends, while the kinetochore
microtubules are being shortened by the loss of tubulin subunits from their
depolymerising ends.
iii) Interaction of Maotubules with other Components
A number of proteins other than tubulin have recently been detected within the
mitotic spindle. The components other than microtubules may provide the driving
force for chromosome movement. Proteins like actin and myosin which have been
detected in close association with the spindle apparatus suggest their possible role in
chromosome movement. The role of actin andjmyosin in other types of motility is
well established. However, their role in chromosome movement has not been
established as yet.
 Ceu Cycle .ad Mitosis

Let us summarise what we have learnt so far.

A growing cell consists of two essential periods, interphase i.e. the period of non-
apparent division and the period of division. The cell cycle is the complex series of
stages by which cellular material is divided equally between two daughter cells.

Doubling of DNA takes place during interphase and the synthesis of DNA occurs
only in restricted portions of the interphase called S-period followed by the "gap"
periods of interphase (GI and G2) in which there is no DNA synthesis. S-phase
cells contain a factor which induces DNA synthesis and this phase lasts for several
hours. The regulation of the cell cycle occurs in the G1 phase, during which it has
to decide whether the cell will start a new cell cycle or become arrested in the GO.
state. Once this G1 check point has been passed, the cell goes on to complete a
new cycle.

Mitosis is a complex process that splits chromosomes equally into two daughter
cells and maintains chromosomal continuity and diploid number. The stage of
mitosis comprises interphase, early prophase, middle prophase, late prophase,
metaphase, early anaphase, late anaphase, telophase and cytokinesis.

Assembly of spindle is regulated by the polymerised and unpolymerised states of

tubulin. During interphase, centrioles are surrounded by the scattered cluster of
short microtubules. At the beginning of prophase, the two pairs of centrioles and
their associated aster begin to separate and move towards opposite poles of the
cell. By the time, centrioles have reached opposite sides of the cell, the spindle
microtubules almost completely disappear. Microtubules exhibit an intrinsic
polarity, one $nd functioning in tubulin assembly and the other functioning in
disassembly.

Two distinct types of movements occur during anaphase i.e. migration of the
chromosomes towaras the poles and the movement of the two poles away f r o r ~
one another. Spindle microtubules play a central role in these two types of
movementc
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16.6 TERMINAL Q L . T I O N S
1) What are cell cycle and period of intense biosynthetic activity?
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Cell Division, Cell 2) Describe the factors that regulate cell division.
Movement and
Differentiated Cell Types ..............................................................................................................................................
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3) State the role played by 'spindle in mitotic cell division:
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16.7 ANSWERS
setf--tQnestions
1) (i) True (ii) False (iii) False (iv) True (v) False (vi) False (vii) True
2) a) (i) Corfect, b) (ii) Correct, c) (i) Correct, d) (iii) Correct
3) i) The beginning of the cell division is signalled by the appearance of
chromosomes as thin threads inside the nucleus.
ii) During telophase nuclear fragments begin to form nuclear-membrane.
iii) Assembly of spindle is regulated by changes in equilibrium between the
polymerised and unpolymerised states of tubulin.
iv) Kinetochore microtubules are the'extemd structures attached to the moving
chromosomes and the chromosomes movemeny can be inhibited by
exposing cells to microtubule disrupting agents such as colchicine.

Terminal Qnestions
1) A growing cell undergoes a cell cycle which comprises essentially two periods
(1) the period of non-apparent division and (2) the period of division. The
interphase is a period of intense biosynthetic activity in which the cell doubles in
sizs and duplicates precisely its chromosome complement.
2) Certain direct evidences indicate that conditions such as lack of nutrients or
appropriate growth factors limit cell growth. Cytoplasmic factors present in
S-phase are known to be capable of triggering the premature entry of G1 nuclei
' into S-phase.
3) In eucaryotic cells, chromosome movement is dependent on the presence of the
mitotic spindle. Spindle formation is regulated by changes in the equilibrium
between the polymerised and unpolymerised states of tubulin. Several
observations suggest that the spindle microtubules play a central role in the
movement of the two sets of chromosomes to opposite poles of the spindle
during anaphase.