Cell Division

Essential idea: Cell division is essential but must be controlled

Nature of Science: Serendipity and scientific discoveries - the discovery of cyclins was
accidental

Understandings :
- Mitosis is division of the nucleus into 2 genetically identical daughter cells
- Chromosomes condense by supercoiling during mitosis
- Cytokinesis occurs after mitosis and is different in plant and animal cells (mainly due to
the presence of cell wall in a plant cell)
- Interphase is a very active phase of the cell cycle with many processes occurring in the
nucleus and cytoplasm
- Cyclins are involved in the control of the cell cycle
- Mutagens, oncogenes and metastasis are involved in the development of primary and
secondary tumours.

The role of mitosis:

- Mitosis is the division of the nucleus into 2 genetically identical daughter nuclei
- Mitosis can only happen in eukaryotic as it involves the division of nucleus
- Replication of DNA prior to mitosis ensures the doubling of DNA molecules forming
chromatids. This is to distribute the number of chromosomes evenly between the 2
daughter cells.
- Before cell division, each chromosome is a single layer. After cell division, it becomes a
double layer, and each of these double layers is called a chromatid.
- Mitosis is involved in growth, tissue repair, tissue replacement, development of embryo,
asexual reproduction and initial stages of gametogenisis
- Growth: In sexual reproduction, a zygote is formed and from the zygote a
multicellular organism is formed. To form this organism, the Zygote needs to go
through repeated cell division which is mitosis
- Tissue Repair: all wounds heal. The cells divide through mitosis and repair the
damaged tissue, and hence heal the wounded region
- Tissue replacement: sometimes some cells in the tissue die and need to be
replaced. Fresh cells in the tissue are produced through mitosis. Example: RBC.
They have a very limited lifespan of 120 days. Fresh RBC need to be produced
via mitosis
- Asexual reproducution: similar to binary fission.
- Gametogenesis. This is the production of gametes. Later on it is meiosis, but
initially it is mitosis.
Cell cycle in Eukaryotes:
- The cell cycle consists of Interphase and M Phase
- Cytokinesis: Division of nucleus
- Karyokinesis: Division of cytoplasm
- G1 : Growth 1
- S : Synthesis
- G2: Growth 2
- In order for cells to survive, they need to maintain
specific surface area/ volume ratio, hence they
divide
- Generally this cell cycle in 24 hours, but sometimes
it can be more or less
- Sometimes the cells may remain in interphase permanently, which means the cell is not
able to divide. Example: neurons, cardiac muscles

Interphase: divisible into 3 stages - G1, S, and G2 phase

G1 phase:
- Synthesis of RNA
- This synthesis is from DNA. The DNA acts like a template. This has to happen
because all the information present in DNA has to be translated into proteins.
Proteins specifically synthesize in the ribosomes.
- Synthesis of certain proteins
- Chromosome is the form of a single strand:
- 2 arms held together by a centromere
- (from DNA the information is passed on to RNA. The RNA, being smaller in size, can
leave the nucleus through the nuclear pore and reach the cytoplasm. In the cytoplasm,
all the information in the RNA becomes translated into proteins)
- (DNA → RNA → Proteins)
S phase:
- DNA synthesis/replication
- DNA has a double helix structure. From a
single DNA molecule, 2 dotted DNA
molecules are produced. This process is
called Replication.
- This is needed for the single stranded
chromosome to become a double
stranded chromosome.
- Each strand is now called a chromatid
- Duplication of centrioles in animal cells
- (For DNA, it replication, for centrioles, its
duplication)

G2 phase:
- More RNA and protein synthesis
- This is mainly needed to increase the cell size
- Duplication of cell organelles
- multiplied by 2 such as mitochondria
- Cell cytoplasm increases, and the cell grows in size
- It distributes the cell organelles evenly to the 2 daughter cells)
- (This is preparation for division.)

M Phase: Divisible into 2 stages:

- Karyokinesis : Division of nucleus. Eukaryotic diploid nucleus divides into
2 diploid daughter nuclei
- The total number of chromosomes in diploid cells is described as 2n
- This is why mitosis is also known as equational division
- Further divided into 4 stages:
- Prophase
- Metaphase
- Anaphase
- Telophase
- Cytokinesis: Division of cytoplasm that divides entire cell into 2 daughter cells each
enclosing a diploid nucleus
 DNA Packaging and Supercoiling of chromosomes:

- The thin strand of DNA(2nm) goes under supercoiling

- A diploid cell contains about 2 meter long DNA when
stretched (146 base pairs)
- Gets packed into the cell nucleus of 5-100 micron in
diameter
- The coiling process happens because of condensation.
- When a piece of chromosome is taken, you can see
supercoiling
- When a piece of the supercoiling is taken, there are
scaffold proteins, which indicates coiling within supercoiling
- When you take a small portion of the scaffolding protein,
you can find looped domains, called a chromatin fibre.
- When a piece of chromatin is enlarged, you can see
histone and DNA, which shows the double helix of DNA.
- Surrounding the 8 histone proteins is the DNA molecule
- Nucleosome: 8 histone proteins surrounded by the DNA.
Together this forms a nucleosome
- DNA with nucleosome is now 11nm
- Chromatosome: 8 histone proteins, the DNA molecule, and
H1 histone protein form a chromatosome

Definitions:
- Centriole: A centriole is the main component of the centrosome. It is made up of
microtubules
 - Centrosome: It is an organelle located near the nucleus in the cytoplasm that divides
and migrates to opposite poles of the cell during mitosis. Centrosome typically contains
two centrioles.
- Centromere: the point or region on a chromosome to which the spindle attaches during
mitosis and meiosis.
- Chromatin: Chromatin is a substance within a chromosome consisting of DNA and
histone protein.
- Chromatid: A chro matid is one of two identical paired strands of a replicated
chromosome joined by a single centromere.
- Chromatosome: Chromatosomes are fundamental units of chromatin structure that are
formed when a linker histone protein binds to a nucleosome
- Chromosome: a threadlike structure of nucleic acids and protein found in the nucleus of
most living cells, carrying genetic information in the form of genes. Each cell normally
contains 23 pairs of chromosomes.

DNA 2nm

Nucleosome 11nm

Solenoid 30 nm

Looped Domain 300 nm

Chromatid 700 nm

Chromosome 1400 nm
Mitosis:
- equational division : no change in chromosome number (same numbers parent)
- diploid number of chromosomes is needed: cannot be an odd number (prophase in this
diagram is wrong it cannot be 3)
- Prophase, Metaphase, Anaphase, Telophase
Prophase:
- It begins as soon as interphase is over

- The long, thin, thread-like chromosomes coils to become thick and

short- this process is called condensation of chromosomes
- As prophase continues, condensation continues and towards the end of this
chromosomes are clearly visible
- Each chromosome is composed of 2 chromatids held
together in a region called centromere
- Polar migration of centrioles begins in prophase: out of 2
pairs of centrioles, one pair begins to move away to the
opposite pole
- Microtubules surround each pair of centrioles
- Delicate fibrils appear around each pair and each looks like a
star. Hence it is called an aster. Since plant cells do not have
centrioles, they do not produce sters.
- Some fibrils extend from one aster to the other, producing
spindle fibres (spindle fibres are made from microtubules)
- This spindle fibre formation is triggered by all the metabolic
actions happening
- Asters are not found in plant cells as there are no centrioles.
The spindle fibres are in astral
- Animal cells the spindle fibres are astrous
- The nuclear envelope breaks down and distingerates
- Nucleolus shrinks in size and disintegrates
Metaphase:
- Since the nucleus has disappeared, the chromosomes have been set free
into the cytoplasm
- As seen in prophase, there is one set of spindle fibres that are shorter and
another that extend towards each other
- There are also another pair of spindle fibres that are intermediate in length,
between the astral spindle fibres and polar spindle fibres
- These spindle fibres become attached to the centromeres of the
chromosomes
- All the chromosomes are arranged with their centromeres
on the equator of the spindle-metaphase plate
- This arrangement is aided by the spindle fibres
- They arrange themselves in such a way that they
centromeres lie on the equator and the chromatids are set
free in the cytoplasm
- Each centromere is attached to the spindle fibres
- Kinetochore: There are special proteins located here. Only
in this region can the spindle fibres attach.
-
- Now all the spindle fibres constitute spindle apparatus : first
type surrounds the aster, the second type are attached to
the centromere of chromosomes , and the third typenstarts
at one pole and extends to the other pole
- The spindle apparatus is astral
Anaphase:
- The spindle fibres contract and divide the centromeres
- In some cases, the spindle fibres contract, or they disassemble from the polar region.
When it contracts, it pulls the centromere towards the pole
- Now from the polar ends, since its fixed, it pulls the centromere to the opposite end
- As a result, the centromere breaks in the center
- Now half of the centromere with one chromatid goes to one pole and the second
chromatid goes to the other pole
- Now at anaphase, the single chromatid
itself is consider a chromosome
- Hence there is no change in the
number of chromosome, just now
instead of 2 chromatids it is a single
chromatid
- Hence the chromatids of each
chromosomes separate and are now
called “daughter chromosomes”
- The daughter chromosomes move to the opposite poles
- Cytokinesis begins towards the end of anaphase

Telophase:
- The daughter chromosomes reach the opposite poles. Their appearance changes
- They uncoil, hence become thin and long once again
- The spindle fibres break down and the spindle fibres disappear
- A new nuclear envelope is formed around each set of chromosomes forming nuclei
- The nucleolus has started to reappear
- Telophase is said to be the opposite of prophase
- Karyokinesis has completed
- Cytokinesis has not completed yet
Cytokinesis:
- Begins in late anaphase
- Cytokinesis is centripetal in case of animal cells
- In animal cells, it takes place by a process called furrowing
- A cleavage furrow develop at the equatorial surface of the cell membrane
- The constriction deepens further, till the center of the cell, dividing the cell into 2
daughter cells
- Cytokinesis is centrifugal in case of plant cells
- The plant cell is covered by a rigid cell wall and the process of cytokinesis cannot occur
by the constriction of the cell membrane
- In case of plant cell, cytokinesis starts in the center and moves along towards the
periphery
- Small vesicles appear at the center of the cell at late anaphase, called a cell plate
- These vesicles grow outwards, till the periphery and form the cell wall
- Once the cell plate has formed, on either side of the cell plate, other vesicles containing
cell wall material other than cellulose like peptin etc, they adhere to the cell plate forming
the middle lamella
- There is not a complete separation yet in plant cells
Mitosis - photomicrographs
From top left to bottom right:
- The chromosomes condense and appear as long
thread-like structure - prophase
- They then align along the center of the cell
(metaphase)
- The micrograph has a darker stain color which
are the chromosomes
- Each chromosome consists of identical sister
chromatids that separate and are pulled to
opposite ends of the cell - anaphase
- Nuclear membranes then form around the 2
daughter nuclei and the chromosomes
decondense.

TEM of human lymphocyte: polar view

Control of cell cycle:
- Cyclins are a family of regulatory proteins that control
the progression of the cell cycle
- Can happen at any stage (G1, M, G2)
- Cyclins activate cyclin dependent kinases (CDKs),
which control cell cycle processes through
phosphorylation (addition of phosphate group)
- CDK has a binding site for cyclin
- When a cyclin and CDK form a complex, the complex
will bind to a target protein and modify it via
phosphorylation
- To convert target protein from inactive stage to active
stage, a cyclin is required
- The phosphorylated target protein will trigger some
specific event within the cell cycle (example:
centrosome duplication, etc)
- After the event has occurred, the cyclin is degraded
and the CDK is rendered inactive again
- Enzymes present in the cell are utilized to break down
the cylin
- The level of cyclin varies in different stages of mitosis, based on internal or
environmental factors
- If there are not sufficient target proteins, there will be no progression of cell cycle

Different Types of Cyclin:

- Cyclin D triggers cells to move from G0(resting phase) to G1 and from G1 to S phase.
- It monitor the content of the cell but also the condition of the cell, whether it is
ready for cell division or not
- Cyclin E prepares the cell for DNA replication in S phase
- All the raw material required for DNA replication is accumulated in the nucleus
- Cyclin A activates DNA replication inside the nucleus in S phase
- Cyclin B promotes the assembly of the mitotic spindle and other tasks in the cytoplasm
to prepare for mitosis
- It also makes sure that the spindle fibres get attached to the kinetochore. At least 2
spindle fibres per kinetochore
Cyclin Expression Cycle:
- Cyclin D gradually increases and then gradually decreases.
- Cyclin E : DNA replication hence a very short duration of time
- Cyclin A : throughout S phase it keeps increasing and in G2 phase there is no DNA
replication hence it drops
- Cyclin B: it gradually starts in G1 and in early metaphase, spindle fibres appear and then
they contract and disappear, which is why there is a sudden drop
- Any of these cyclins can prevent the progress of cell division

G1 phase checkpoint:
- It determines whether all conditions are favorable for cell division to proceed
- It is regulated.
- Cyclin D can bind with 2 different types of CDK: CDK4 and CDK6. It regulates
pre-requisite conditions required for the cell. Cyclin D is needed to maintain the
conditions needed for cell division. Only then will it allow the cyclin to pass from G1 to S
phase.
- If any of the conditions are not met or any proteins are nor produced, cyclin D prevents
the cell from entering S phase. The cell can half the cycle and attempt to remedy the
problematic condition, or the cell can advance into G0 and await further signals when
conditions improve

S phase checkpoint:
- for S phase to progress, 2 cyclins are needed : A and E
- cyclin E prepares the entire cell for DNA replication in which the parent DNA splits into 2
daughter DNA.
- Cyclin A is responsible for the nucleus. cyclin A is not there, there is no DNA replication.
- One prepares the cell and the other initiates the process.
- If either cyclin is in lower level, there will be no progression from S phase to G2 phase.

G2 phase checkpoint:
- The G2 checkpoint bars entry into the mitotic phase if certain conditions are not met.
- As at the G1 checkpoint, cell size and protein reserves are assessed.
- However, the most important role of the G2 checkpoint is to ensure that all of the
chromosomes have been replicated and that the replicated DNA is not damaged
- Cyclin A is needed as if it is not there, no further protein synthesis.
- If the checkpoint mechanisms detect problems with the DNA, the cell cycle is halted, and
the cell attempts to either complete DNA replication or repair the damaged DNA\

M checkpoint:
- Occurs near the end of metaphase
- Also known as spindle checkpoint because it determines whether all the sister
chromatids are correctly attached to the spindle microtubules
 - Because the separation of the sister chromatids during anaphase is an irreversible step,
the cycle will not process until the kinetochores of each pair of sister chromatids are
firmly anchored to at least 2 spindle fibres arising from opposite poles of the cell
- regulated by cyclin B. if there is no cyclin B, there is no check on spindle fibres, getting
attached to the kinetochore, so it will not progress.

Cancer and cell cycle:

- All regulators are positive regulators.
- There needs to be a gene responsible for CDK protein synthesis
- Benign tumours: those which can be easily surgically removed
- Cancer comprises many different diseases caused by a common mechanism:
uncontrolled cell growth
- Even when all of the cell cycle controls are fully functional, a small percentage of
replication error (mutations) will be passed on to the daughter cells.
- The genes that code for the positive cell cycle regulators are called proto-oncogenes.
Proto-oncogenes are normal genes that, when mutated in certain ways, becomes
oncogenes, genes that cause a cell to become cancerous
- Many reasons for mutation, for example exposure to radiation (physical, due to certain
chemicals (cigarette smoke), biological mutagens(bacteria or viruses)
- 2 groups of proteins, called cyclins and cyclin-dependent kinases (CDKs) are
responsible for the progress of the cell through the various checkpoint
- The CDK gene is one of the many genes that are considered proto-oncogenes In
addition to the cell cycle regulatory proteins, any protein that influences the cycle can be
altered in such a way as to override cell cycle checkpoints. An oncogene is any gene
that, when altered, leads to an increase in the rate of cell progression
Negative regulators of cell cycle:
- The second group of cell cycle regulatory
molecules are negative regulators.
Negative regulators halt the cell cycle
- The best understood negative regulatory
molecules are retinoblastoma protein (Rb),
p53, and p21. They act primarily at the G1
checkpoint
- Retinoblastoma proteins are a group of
tumour-suppressor proteins common in
many cells
- (P stands for protein, the number(ex: 53,
21) stands for the atomic mass in daltons)
- P53 is a multi-functional protein. During the
S phase when DNA synthesis is done, it checks the conditions. if DNA damage halts the
cell cycle and it triggers DNA repair. If the DNA cannot be repairs, p53 can trigger
apoptosis, or cell suicide, to prevent the duplication of damaged chromosomes. If this is
not done it could lead to abnormality.
- As p53 levels rise, the production of p21 is triggered. P21 enforces the halt on the cycle
dictated by p53 by binding to and inhibiting the activity of the CDK/cyclin complexes
- As a cell is exposed to more stress, high levels of p53 and p21 accumulate, making it
less likely that the cell will move into the S phase
- For the cell to move past each of the checkpoints, all positive regulators must be “turned
on” and all negative regulators must be “turned off”

Telomere and cancer:

- Telomere is at the tip of each
chromosome, and protects the
chromosome from the action of some of
the enzymes
- In a normal cell, as the cell ages the
telomere length decreases, it also affects
cell division. The cell loses its ability to
divide. Now we say the cell has aged and
ultimately leads to the death of the cell
- Unlike in a normal cell, once cancer cells
get telomerase on, they never turn it off
- Instead the enzyme just keeps adding
more and more repeats to the telomeres
- Now the cancer cell can keep dividing
without losing DNA and genes at the ends of the chromosomes
- Metastasis will occur, where the cancer spreads to the rest of the body
- Chemotherapy uses chemicals to prevent the growth of the cell. They may contain
telomerase enzyme inhibitor
 - Telomerase inhibitors are useful to control the multiplication of the cancerous cells. It
prevents the action of telomerase enzyme, so the telomere will stop growing and stops
dividing.

Mitotic Index:
- Used to see whether the cells are actively dividing and to see the response of the
cancerous cell after chemotherapy
- It can be calculated using this formula
- Page 57 and 58 of TB