Nuclear

Structure and
Function
PHRA 312
Chap4

Sreedam Das, PhD II Associate Professor

Dept. of Clinical Pharmacy and Pharmacology
University of Dhaka sreedamspph@iub.edu.bd
 Life starts as
a single cell

37.2 Trillion cells Cell Division

The way to grow, develop
and repair our body

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 Cell cycle
▪ Cell cycle is an ordered series of events that lead to cell division and the
production of two daughter cells each containing chromosomes identical to those
of the parental cell.

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 Cell cycle

Interphase Mitosis
Prophase
G1 (gap 1) phase
Prometaphase
S (synthetic) phase Metaphase
G2 (gap 2) phase Anaphase
Telophase

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 G1 phase

▪ The first phase of interphase.

▪ The cell usually grows larger, and some organelles are copied.
▪ Cells take preparation to replicate DNA by synthesising the mRNAs and
proteins required to execute the future steps.

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S phase
▪ All the genetic information (DNA) in the cell is copied by the process of DNA
replication.
▪ DNA replication is semi conservative and semi discontinuous type which results in
the formation of identical pairs of DNA molecules.
▪ DNA replication generates sister chromatids, which are identical pairs of
chromosomes. So after replication, number of chromosome is still 46 but cell
contains 92 chromatids. (The number of chromosome = the number of functional
centromere. The number of DNA molecule= the number of chromatids)
▪ These sister chromatids are attached to each other by a centromere.
▪ The centrosome (centriole) is also duplicated during the S phase.

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 G2 phase

▪ The final phase of interphase is the G2 phase.

▪ Cell grows further, makes proteins and organelles.
▪ Metabolic changes assemble the cytoplasmic materials necessary for
mitosis.​
▪ The phase ends when mitosis begins.

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 Mitosis (M)

▪ Mitosis is a form of eukaryotic cell division that produces two daughter

cells with the same genetic component as the parent cell.
▪ Chromosomes replicated during the S phase are divided in such a way as
to ensure that each daughter cell receives a copy of every chromosome.
▪ In actively dividing animal cells, the whole process takes about one hour.

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 Mitosis (M)
▪ Mitosis can be divided into five phases:
o Prophase
o Prometaphase
o Metaphase
o Anaphase
o Telophase
▪ Sometimes, prometaphase is not considered a separate phase.

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(i) Prophase
▪ Prophase is the first stage of cell division in both mitosis and meiosis.
▪ It begins after interphase where DNA has already been
replicated (duplicated with identical copies of sister
chromatids; 46 chromosome with 92 chromatids ).
▪ The chromatin begin to coil and compact, resulting in the
formation of visible chromosomes of X shape.
▪ Centrosomes (duplicated in S-phase) start to move to
opposite poles of cells and spindle fibres begin to form.
▪ Spindle fibre (microtubules) begins to form
▪ Nucleolus disappears

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 (ii) Prometaphase
▪ Prometaphase is not always presented as a distinct
part of mitosis. It is the phase to late prophase and
early metaphase.

▪ Nuclear membrane breaks apart into numerous

"membrane vesicles”
▪ The kinetochore assembles on the centromere and
links the chromosome to microtubule

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(iii) Metaphase
▪ The centromeres of the chromosomes convene
themselves on the metaphase plate, an imaginary line
that is equidistant from the two centrosome poles.
▪ The even alignment at metaphase plate is due to the
counterbalance of the pulling powers generated by
the opposing kinetochore microtubules.
▪ Unattached or improperly attached kinetochores (if any) generate a signal to prevent
premature progression to anaphase, even if most of kinetochores have been
attached and most of the chromosomes have been aligned. Such a signal creates the
mitotic spindle checkpoint.
▪ Metaphase is the crucial part in cell division as all of the genetic material should be
perfectly connected to kinetochore to ensure exact copy of chromatid moves to
opposite poles.
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(iv) Anaphase

▪ Anaphase is the stage after metaphase, when

kinetochore-connected chromatids are moved
to opposite poles of the cell.
▪ Anaphase starts when the ‘anaphase
promoting complex’ destroys the enzyme
known as ‘Securin’.
▪ The destruction of Securin unleashes the
protease enzyme ‘Separase’.
▪ Inactivation of of Seperase causes breakdown
of ‘Cohesin’, a protein responsible for holding
sister chromatids together.
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 (v) Telophase
▪ Telophase is the final stage in both meiosis
and mitosis
▪ At this stage, the nucleolus reappear and
nuclear membrane is reassembled (during
prophase and prometaphase it was
disintegrated into membrane vesicles).

▪ Chromosomes begin to decondense back into the expanded chromatin that is

present during interphase.
▪ The mitotic spindle is disassembled and remaining spindle microtubules are
depolymerized.

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Cytokinesis

▪ Cytokinesis completes the cell cycle,

and usually overlaps with the final
stage (telophase) of mitosis.
▪ Cytokinesis involves in the physical separation of the cytoplasm and its components
into daughter cells.
▪ A cleavage furrow separates the daughter cells
▪ Some eukaryotic cells may omit cytokinesis producing multinucleated cells. For
example: hepatocytes and skeletal muscle cells.

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 This is due to the process of Meiosis
where genetic diversity happens during generation of the
gametes for reproduction

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 Meiosis

▪ Meiosis a special type of cell division to produce sex cells or gametes, such as
sperm in males and egg in females.
▪ In this process haploid cells are produced from a diploid cells.
▪ Following DNA replication (duplication of 46 chromosome to produce 92
chromatids in S phase of cell cycle) meiosis causes two rounds of cell division to
produce four daughter haploid cells.
▪ The goal of meiosis is to make daughter cells with exactly half of chromosomes of
the starting cell so that after fertilization from sperm and eggs to create a cell with
two copies of each chromosome again.

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 Meiosis
▪ Meiosis I and II are each divided into prophase, prometaphase, metaphase,
anaphase, and telophase stages, similar in purpose to their analogous subphases in
the mitotic cell cycle.
▪ Therefore, meiosis includes the stages of meiosis I (prophase I, prometaphase I,
metaphase I, anaphase I, telophase I) and meiosis II (prophase II, prometaphase II,
metaphase II, anaphase II, telophase II).

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 DNA replication at S phase of cell cycle

DNA (DNA replication in S phase produces two identical copies of 46 chromosomes

with 92 chromatins) condenses into X-shaped structures that can be easily seen
under a microscope.

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Prophase I
▪ Prophase is the first stage of cell division in both mitosis and meiosis.

▪ It begins after interphase where DNA has already been

replicated (duplicated with identical copies of sister
chromatids; 46 chromosome with 92 chromatids ).
▪ The chromatin begin to coil and compact, resulting in the
formation of visible chromosomes of X shape.
▪ Centrosomes (duplicated in S-phase) start to move to
opposite poles of cells and spindle fibres begin to form.
▪ Spindle fibre (microtubules) begins to form
▪ Nucleolus disappears
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Prophase I

▪ The chromosomes pair up so that both copies of chromosome 1 are together, both
copies of chromosome 2 are together, and so on.

▪ Homologous pairs of sister chromatids lies side

by side in a process called synapsis forming a
tetrads or bivalents.
▪ The bivalents or tetrads have two homologous
chromosomes and four chromatids, (somatic
cells have two sets of chromosomes, one set
coming from mother and another set from
father. So 23 pair of homologous chromosomes
with 92 chromatids).

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 ▪ Once bivalent is formed, the second event called crossing over happens.
▪ The site where cross-over happens is called chiasmata (plural; singular
chiasma)
▪ During crossing over, a physical exchange between chromosome segments of
non-sister chromatid occurs, thus increasing genetic diversity.
▪ Non-sister chromatids have similar not necessarily identical genes. The
crossing over in non-sister chromatids thus causes genetic diversity in haploid
sex cells.

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 Prometaphase I
▪ It is the phase to late prophase and early metaphase.
▪ Nuclear membrane breaks apart into numerous
"membrane vesicles”
▪ The kinetochore assembles on the centromere and
links the chromosome to microtubule

▪ Here is a key difference between meiosis and mitosis prometaphase.

o In mitosis, EACH CHROMATID is attached to different fibres from opposite
poles. In meiosis, each PAIRS OF SISTER CHROMATIDS (two identical
chromosomes) are attached to mitotic fibre from opposite poles. This leads to
the formation of haploid cells.

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Prometaphase I

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Metaphase I
▪ The bivalent align along the metaphase plate, an
imaginary line that is equidistant from the two
centrosome poles.
▪ The even alignment at metaphase plate is due to
the counterbalance of the pulling powers generated
by the opposing kinetochore microtubules.
▪ During this alignment, independent assortment
happens which leads to another genetic diversity.

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Anaphase I
▪ Anaphase is the stage after metaphase,
when kinetochore-connected
chromosomes (with sister chromatids) are
moved to opposite poles of the cell.
▪ In anaphase I the sister chromatids remain
attached at their centromeres and move
together toward the poles
▪ So, 23 chromosomes (46 chromatids) move
to two opposite poles.

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Telophase I
▪ During this phase, at each pole, there is
a complete haploid set of chromosomes
(23 chromosomes with 46 chromatids).
▪ Nuclear envelop temporarily arrives.

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Cytokinesis

▪ Cytokinesis involves in the physical

separation of the cytoplasm and its
components into daughter cells.
▪ A cleavage furrow separates the
daughter cells.
▪ Thus cytokinesis from meiosis I produces
two haploid daughter cells with genetic
diversity.
▪ After cytokinesis, the daughter cells
begin immediately to prepare for the
second meiotic division (meiosis-II).

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 Meiosis II

▪ Meiosis II begins without any DNA replication.

▪ Next five steps are:
i. Prophase II
ii. Prometaphase II
iii. Metaphase II
iv. Anaphase II
v. Telophase II

▪ Meiosis II proceeds same cell division steps just like Mitosis.

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 Meiosis II

▪ During anaphase II, kinetochore-connected chromatids are moved to

opposite poles of the cell. [During anaphase I, chromosomes with sister
chromatids moved towards opposite pole].
▪ In telophase II, each daughter cells get 23 chromosomes (23 chromatids).
[During telophase each daughter cells get 23 chromosomes (46 chromatids).

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 After fertilization, cell gets 23 pairs of chromosomes

▪ During fertilization, a sperm is fused with egg forming zygote which grow
into a child.
▪ The child receives half of it chromosome from mother (haploid ovum) and
rest half from father (haploid sperm).
▪ Men and women produce millions of gametes and selection of gametes in
fertilization is random, this is tricky in genetic diversity.
▪ This explains why a child is not identical to either parents.

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Meiosis leads to genetic diversity
Two events happen in meiosis that do not happen in
mitosis and lead to genetic diversity.
▪ Exchange of random genetic information during Prophase I:
o During prophase I of meiosis-I, homologous chromosomes pair up and
exchange random genetic information.
o This results in a new combination of genes in each chromosome. These
altered chromosomes give rise to what are known as "recombinant
chromosomes" in the gametes.
o Because chromosomes may contain hundreds of genes, a single crossover
event can affect many genes.
o Since more than one crossover can occur in each tetrad, it is no wonder that
gametes and the offspring that result from them can be so varied.
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 Exchange of random genetic information during Prophase I

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Meiosis leads to genetic diversity
▪ Independent assortment during Metaphase-I:
o The chromosomes in each homologous pair (tetrads) line up and separate at
metaphase I is a matter of chance.
o The assortment of chromosomes that end up in the resulting cells occurs
randomly.
o If we consider a diploid cell with four chromosomes (two homologous pairs),
there are two equally possible ways for the chromosomes inherited from the
two parents to be arranged during metaphase I. This variation in the
orientation of chromosomes leads to gametes with four equally possible
combinations of chromosomes (Figure-next page).

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 For the organism in this Figure, n = 2, so the number of chromosome combinations is 22, or 4.
For a human, n = 23, so there are 223, or about 838,860 possible chromosome combinations!

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Meiosis leads to genetic diversity
▪ Fertilization
o Fertilization presents yet another opportunity for genetic diversity.
o Millions of genetically different sperm swimming toward an egg. Fertilization is
random, so the sperm that wins the race in one fertilization event is going to be
different than the sperm that wins the next race. And, of course, each egg is
genetically different too.
o So, fertilization produces random combinations of genetically diverse sperm and
eggs, creating virtually unlimited possibilities for variation.
o That’s why every human being is genetically unique. Genetically identical twins
can develop from the same fertilized egg, but even they can have subtle
differences due to development.

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 Cell cycle needs to be highly regulated and controlled fashion

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 Regulation of the cell cycle

▪ During cell division, it is essential that the daughter cells exactly duplicate the
parent cell.
▪ To prevent a compromised cell from continuing to divide, internal control
mechanisms have developed cell cycle checkpoints.
▪ Cell cycle checkpoints are control mechanisms in the eukaryotic cell cycle which
ensure its proper progression.
▪ Each checkpoint serves as a potential termination point along the cell cycle,
during which the conditions of the cell are assessed, with progression through the
various phases of the cell cycle occurring only when favorable conditions are met.

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 Regulation of the cell cycle

▪ There are four checkpoints in the cell cycle:

o G1 checkpoint/ start or restriction checkpoint/major checkpoint
o S checkpoint
o G2 checkpoint
o M checkpoint/ mitotic checkpoint/ spindle checkpoint

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Regulation of the cell cycle

▪ There are multiple CDK-cyclin complexes (CDKCs) that play specific roles at various
phases in the cell cycle.
▪ While levels of CDKs remain fairly constant
throughout the cell cycle, cyclin levels fluctuate.
▪ The fluctuation controls the activation of the
CDKCs and ultimately the progression
throughout the cycle.
▪ Specific cyclins cause activation of specific Cdks.

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 The DNA in our cells is packaged into 46 chromosomes
as supercoiled structure.

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 How DNA is packed inside a nucleus?

▪ Packing of our DNA with the assembly

of NUCLEOSOME which consists of a
segment of DNA wound around eight
histone proteins
▪ Nucleosome resembles thread
wrapped around a spool and the
appearance is known as ‘beads on a
string’.
▪ The nucleosome core particle consists
of approximately 146 base pairs of
DNA wrapped around a histone
octamer, consisting of 2 copies each of
the core histones H2A, H2B, H3, and H4.
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 ▪ At the entry and the exit of the nucleosome, histone H1 binds the DNA
to make the nucleosomes fixate in the space.
▪ Nucleosomes are connected by stretches of linker DNA (10-60 bp) and
coil to form an array along the DNA that is about 11 nm in diameter.
▪ Linker DNA contains target sites for one or more restriction enzymes.
The linkers can be synthesized chemically and can be ligated to foreign
DNA or vector DNA containing more restriction sites.

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▪ Multiple nucleosome coiled together, which is then stacked on each other. The end
result is a fibre of packed nucleosome known as CHROMATIN. The fibre to at this
point get thickness of 30 nm.
▪ Chromatin fibre is further lopped and packed to form a SOLENOID which gives a
thickness of 300 nm.
▪ The solenoid structure of chromatin helps to make 40 times smaller of the
compacted DNA.
▪ The solenoid structure then again supercoiled into a CHROMOSOME.
Chromosomes are visible in the light microscope only during mitosis, when they
become highly condensed.
▪ This remarkable folding allows six feet of DNA to fit into the nucleus of a single cell.

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 Types of chromosome
On the basis of the position of centromere

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 Telomeres

▪ Telomeres are nucleoprotein structures at the ends of eukaryotic chromosomes.

▪ Telomeres are essential to protect chromosome ends from DNA degradation and
DNA repair activities and they play an important role in chromosome stability.
▪ Telomeres shorten with every cell division owing to the so-called “end replication
problem”.
▪ Telomerase is a ribonucleoprotein DNA polymerase that can elongate telomeres
by de novo addition of TTAGGG repeats onto the chromosome ends, thus
compensating for telomere shortening.

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 Does telomerase has role in ageing or death?

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Does telomerase has role in ageing or death?

▪ Telomerase enzyme is not sufficient in human cells. So, DNA shortening does
happen in our body each time our DNA replicate or cell divides.
▪ In humans, average telomere length declines from about 11 kilobases at birth
to fewer than 4 kilobases in old age, with the average rate of decline being
greater in men than in women.
▪ Telomere shortening is considered one of the hallmarks of aging as short
telomeres are sufficient to cause organismal aging and decreased lifespan.

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 ▪ When telomeres reach a critically short length, they induce a persistent
DNA damage which induces other cellular events, such as cellular
senescence and/or apoptosis, as well as impairs the ability of stem cells to
regenerate tissues.
▪ When they get too short, DNA molecules start to get damaged, the cell
can no longer divide and eventually dies.

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