Cell Replication

Sry Suryani Widjaja

Biochemistry Department
Sept 2022
CHROMOSOME
• Chromosomes are composed of protein and DNA (chromatin)

• The DNA in an individual chromosome is one long molecule, highly

coiled and condensed.
• Genetic information in the DNA is defined by the linear sequences of
bases (A, T, C and G), the genetic code.
• There may be 50–250 million bases in an individual chromosome.
• DNA sequence for a single trait is called a gene.
• The number of chromosomes in human cells is 46; 22 autosomal pairs
(same in both sexes) and 2 sex chromosomes, 2 X chromosomes in
Females and 1 X and 1 Y in males.
• Chromatin is the complex combination of DNA, RNA and protein that makes up
chromosomes inside the nuclei of eukaryotic cells;
• It is divided between heterochromatin (condensed) and euchromatin (extended)
forms.
• The functions of chromatin are to package DNA into a smaller volume to fit into the
cell, to support the DNA to allow mitosis and meiosis, and to serve as a mechanism
to control expression and DNA replication.
• Changes in chromatin structure are affected by chemical modifications of histone
proteins, such as methylation (DNA and proteins) and acetylation (proteins), and by
non-histone DNA binding proteins.
• Chromatin is easily visualised by staining, hence its name, which literally means
coloured, lightened material.
CELL REPLICATION
• MITOSIS
• Mitosis is the phase of the cell cycle where chromosomes in
the nucleus are evenly divided between two cells.
• When the cell division process is complete, two daughter cells with
identical genetic material are produced.
Interphase

• Before a dividing cell enters mitosis, it undergoes a period of growth called interphase.

• About 90 percent of a cell's time in the normal cell cycle may be spent in interphase.
• G1 phase: The period prior to the synthesis of DNA. In this phase, the cell increases in
mass in preparation for cell division. The G1 phase is the first gap phase.
• S phase: The period during which DNA is synthesized. In most cells, there is a narrow
window of time during which DNA is synthesized. The S stands for synthesis.
• G2 phase: The period after DNA synthesis has occurred but prior to the start of prophase.
The cell synthesizes proteins and continues to increase in size. The G2 phase is the second
gap phase.
• In the latter part of interphase, the cell still has nucleoli present.
• The nucleus is bounded by a nuclear envelope and the cell's chromosomes have
duplicated but are in the form of chromatin.
Prophase
• In prophase, the chromatin condenses into discrete chromosomes.
The nuclear envelope breaks down and spindles form at opposite
poles of the cell.
• Prophase (versus interphase) is the first true step of the mitotic
process. During prophase, a number of important changes occur:
• Chromatin fibers become coiled into chromosomes, with each
chromosome having two chromatides joined at a centromere.
• The mitotic spindle, composed of microtubules and proteins, forms in
the cytoplasm.
• The two pairs of centrioles (formed from the replication of one pair in
Interphase) move away from one another toward opposite ends of
the cell due to the lengthening of the ​microtubules that form
between them.
• Polar fibers, which are microtubules that make up the spindle fibers,
reach from each cell pole to the cell's equator.
• The chromosomes begin to migrate toward the cell center.
METAPHASE
• In metaphase, the spindle reaches maturity and the chromosomes align at the
metaphase plate (a plane that is equally distant from the two spindle poles).
• During this phase, a number of changes occur:
• The nuclear membrane disappears completely.
• Polar fibers (microtubules that make up the spindle fibers) continue to extend
from the poles to the center of the cell.
• Chromosomes move randomly until they attach (at their kinetochores) to polar
fibers from both sides of their centromeres.
• Chromosomes align at the metaphase plate at right angles to the spindle poles.
• Chromosomes are held at the metaphase plate by the equal forces of the polar
fibers pushing on the centromeres of the chromosomes.
ANAPHASE
• In anaphase, the paired chromosomes (sister chromatide) separate and begin
moving to opposite ends (poles) of the cell.
• Spindle fibers not connected to chromatids lengthen and elongate the cell.
• At the end of anaphase, each pole contains a complete compilation of
chromosomes.
• During anaphase, the following key changes occur:
• The paired centromeres in each distinct chromosome begin to move apart.​
• Once the paired sister chromatids separate from one another, each is considered a
"full" chromosome. They are referred to as  daughter chromosomes
• Through the spindle apparatus, the daughter chromosomes move to the poles at
opposite ends of the cell.​
• The daughter chromosomes migrate centromere first and the
kinetochore fibers become shorter as the chromosomes near a pole.​
• In preparation for telophase, the two cell poles also move further
apart during the course of anaphase.
• At the end of anaphase each pole contains a complete compilation of
chromosomes.
TELOPHASE
• In telophase, the chromosomes are cordoned off into distinct new nuclei in the emerging daughter
cells.
• The following changes occur:
• The polar fibers continue to lengthen.
• Nuclei begin to form at opposite poles.
• The nuclear envelopes of these nuclei form from remnant pieces of the parent cell's nuclear envelope
and from pieces of the endomembrane system.
• Nucleoli also reappear.
• Chromatin fibers of chromosomes uncoil.
• After these changes, telophase/mitosis is largely complete.
• The genetic contents of one cell have been divided equally into two.
CYTOKINESIS
• Cytokinesis is the division of the cell's cytoplasm.
• It begins prior to the end of mitosis in anaphase and completes
shortly after telophase/mitosis.
• At the end of cytokinesis, two genetically identical daughter cells are
produced.
• These are diploid cells, with each cell containing a full complement of
chromosomes.
Meiosis
• Meiosis is a two-part cell division process in organisms that sexually
reproduce.
• Meiosis produces gametes with one half the number
of chromosomes as the parent cell.
• In some respects, meiosis is very similar to the process of mitosis yet
it is also fundamentally different from mitosis.
• The two stages of meiosis are meiosis I and meiosis II.
• At the end of the meiotic process, four daughter cells are produced.
• Each of the resulting daughter cells has one half of the number of
chromosomes as the parent cell.
• Before a dividing cell enters meiosis, it undergoes a period of growth
called interphase.
• During interphase the cell increases in mass, synthesizes
DNA and protein, and duplicates its chromosomes in preparation for
cell division.
Meiosis I

• Meiosis I encompasses four stages:

• Prophase I - chromosomesa condense and attach to the nuclear envelope
and begin migrating toward the metaphase plate.
• This is the stage where genetic recombination may occur (via crossing
over).
• Metaphase I- chromosomes align at the metaphase plate.
• For homologous chromosomes, the centromeres are positioned toward
opposite poles of the cell.
• Anaphase I - homologous chromosomes separate and move toward
opposite cell poles.
• The sister chromatids remain attached after this move to opposite
poles.
• Telophase I-cytoplasm divides producing two cells with
a haploid number of chromosomes.
• Sister chromatides remain together. While different cell types may
prepare differently for meiosis II, there is one variable that does not
change: the genetic material does not undergo replication in meiosis
II.
Meiosis II

• Meiosis II encompasses four stages:

• Prophase II - chromosomes begin migrating to the metaphase II plate. These
chromosomes do not replicate again.
• Metaphase II - chromosomes align at the metaphase II plate while the
kinetochore fibers of the chromatids are oriented toward opposite poles.
• Anaphase II - sister chromatides separate and begin moving to opposite ends of
the cell. The two cell poles also grow further apart in preparation for telophase II.
• Telophase II - new nuclei form around daughter chromosomes and the cytoplasm
divides and forms two cells in a process known as cytokinesis.
• At the end of meiosis II, four daughter cells are produced. Each of these resulting
daughter cells is haploid.
• Cells produced through mitosis are different from those produced
through meiosis.
• In meiosis, four daughter cells are produced.
• These cells are haploid cells, containing one-half the number of
chromosomes as the original cell. 
• Sex cells undergo meiosis.
• When sex cells unite during fertilization these haploid cells become a
diploid cell.​
•  Errors in meiosis can also lead to genetic disorders.
• One such error is chromosomal non-disjunction.
• With this error, the chromosomes do not separate as they should during
the meiotic process.
• The gametes that are produced do not have the correct number of
chromosomes.
• In humans, for example, a gamete may have an extra chromosome or be
missing a chromosome.
• In such cases, a pregnancy that resulted from such gametes could end in
a miscarriage. 
Chromosome banding

• Chromosome banding is evident when chromosomes are treated with

chemical dyes, such as Giemsa
• chromosomes appear as a series of alternate dark (G-band or G-
positive band) and light (R-band or G-negative band) bands.
• Each chromosome consists of two arms separated by the centromere.
• The long arm and short arm are labelled q (for queue) and p (for petit),
respectively.
• At the lowest resolution, only a few major bands can be distinguished,
which are labelled q1, q2, q3 . . . ; p1, p2, p3 . . . , counting from the
centromere.
• Higher resolution reveals sub-bands, labelled q11, q12, q13 . . . Sub-
sub-bands identified by even higher resolution are labelled q11.1,
q11.2,q11.3 . . .
• Traditionally, the short arm (p) is displayed on top of the long arm (q).
KARYOTYPE
• A karyotype is the representation of the entire metaphase
chromosomes in a cell, arranged in order of size.

• The band width and order of bands is characteristic of a particular

chromosome, and identifiable by a trained cytogeneticist.
• Human somatic cell contains two sets of homologous chromosomes,
which may be divided into two types: autosomes and sex
chromosomes.
• Autosomes are further divided into seven groups: A–G.
The spectral karyotype: fluorescence
in situ hybridisation (FISH)

• So-called ‘chromosome painting’ refers to the hybridisation of fluorescently

labelled chromosome-specific probes.
• Chromosome painting allows the visualization of individual chromosomes in
metaphase or interphase cells, and the identification of both numerical and
structural chromosomal aberrations in human pathology.
• The use of fluorescent dyes that bind to specific regions of chromosomes
can impart unique spectral characteristics.
• Slight variations in colour, undetectable by the human eye, can be quantified
by digital imaging; multicolour (24-colour) fluorescence in situ hybridisation
(mFISH) can provide specific information to improve diagnosis and prognosis
of genetic diseases.
Mendel first law
• Mendel's First Law - the law of segregation; during gamete formation
each member of the allelic pair separates from the other member to
form the genetic constitution of the gamete

• Mendel's First Law states that for the pair of alleles an individual has
of some gene (or at some genetic locus), one is a copy of a randomly
chosen one in the father of the individual, and the other if a copy of a
randomly chosen one in the mother, and that a randomly chosen one
will be copied.
Mendel Second Law: law of independent assortment

• Mendel's 2nd law states that during gamete formation the segregation of

each gene pair is independent of other pairs.
• Mendel's 2nd law is often referred to as the principle of independent
assortment.
• Both of Mendel's laws are about segregation, which is the seperation
of allele pairs.
• The law states that the seperation of one pair of alleles isn't related to the
sepearation of other pairs of alleles, and so is very important in Mendelian
genetics.
• The only time there is an exception to this rule is when linkage is involved.
• There are two types of breeding processes to know the mechanism of
genes and examine the inheritance of traits from parents and
grandparents, one is monohybrid cross and the other is dihybrid
cross.
• A monohybrid cross is the hybrid of two individuals with
homozygous genotypes which result in the opposite phenotype for a
certain genetic trait.”
• “The cross between two monohybrid traits (TT and tt) is called a
Monohybrid Cross.” Monohybrid cross is responsible for the
inheritance of one gene.
• Huntington’s Disease
• Huntington’s diseases is a fatal genetic disorder. The Huntingtin gene,
responsible for Huntington’s disease is present in all the individuals.
• The homozygous dominant Huntingtin gene of an individual was
paired with the homozygous recessive Huntingtin gene of another
individual.
• All the offspring carried the dominant allele for Huntington’s disease.
That means that the child will have this disease.
Dihybrid and Test Cross
• A dihybrid cross is a cross between two different genes that differ in
their observed traits.
• It is a cross between two individuals that are heterozygous for two
different traits.

• Test cross
• a test cross is an experimental cross of an individual organism of
dominant phenotype but unknown genotype and an organism with a
homozygous recessive genotype (and phenotype)
• Back Cross: It is a crossing of a hybrid with one of its parents or an
individual genetically similar to its parent, in order to achieve
offspring with a genetic identity which is closer to that of the parent.
Non Mendelian inheritance
• The eukaryotic cell cycle is composed of four phases.
• Of the four phases, the two critical ones are DNA replication, which occurs
during G1 to S phase, and the physical process of cell division, which
occurs during G2 to M (for mitosis) phase; these are critical ‘checkpoints’.
• In gap phases G1 and G2, the cell is preparing for DNA replication and cell
division respectively.
• M phase is composed of two discrete steps: mitosis, which constitutes the
pairing and separation of the duplicated chromosomes, and cytokinesis,
the physical process whereby the cell splits into two daughter cells.
• Not all cells continue to divide during the life span of an organism;
many undergo what is referred to as ‘terminal differentiation’ and
become quiescent; cells in this phase are said to reside in another gap
phase called G0.

• Under certain conditions, such as that resulting from an external

signal stimulating cell growth, cells can exit the quiescent state and
re-enter the cell cycle.
• Progression through the different phases of a cell cycle is controlled at
‘checkpoints’.
• There are a number of checkpoints, but the two most critical are the
one that occurs near the end of G1, prior to S-phase entry, and the
one near the end of G2, prior to mitosis.
• Cell-cycle control mechanisms is the responsibility of a family of
protein kinases called cyclin dependent kinases (CDKs).
• CDKs are also involved in the regulation of transcription and mRNA
processing.
• A CDK is activated by association with a cyclin, forming a CDK
complex.
• Cyclins are produced or degraded as needed in order to drive the cell
through the different stages of the cell cycle.
Initiation of cell division and
differentiation
• Many cells reside in a resting or quiescent state but can be stimulated by
external signals to re-enter the cell cycle.
• These external growth-promoting signals are the result of growth factors
binding to their receptors.
• Most growth factors induce the expression of genes that are referred to as
early- and delayed-response genes.
• The activation of early-response genes occurs in response to growth factor
receptor-mediated signal transduction, resulting in phosphorylation and
activation of transcription factor proteins that are already present in the cell.
• Many of the induced early-response genes are themselves transcription factors
which in turn activate the expression of delayed-response genes.
• Growth factors and growth factor receptors play an important
physiological role in the normal process of growth and differentiation.
• Genes that are involved in the control of normal cell proliferation can
be classified as proliferative or antiproliferative:
• Proliferative genes are proto-oncogenes; mutated proto-oncogenes
may become oncogenes.
• Antiproliferative or tumour-suppressor genes act to suppress cell
proliferation.
Apoptosis

• Apoptosis, or programmed cell death, is a normal physiological event.

• Apoptosis also functions to remove damaged cells.
• Apoptosis plays a major role in preventing cancer. If a cell is unable to
undergo apoptosis, it continues to divide and develop into a tumour.
• The process of apoptosis is controlled by a diverse range of cell
signals, which may be either extracellular or intracellular.
• Extracellular signals may include toxins, hormones, growth factors,
nitric oxide or cytokines; these must either cross the plasma
membrane or transduce to effect a response
• Two theories for the direct initiation of apoptotic mechanisms in
mammals have been suggested:
• the TNF-induced (tumour necrosis factor) model
• the Fas-Fas ligand-mediated model.
• Both involve receptors of the tumour necrosis factor receptor (TNFR)
family coupled to extrinsic signals.
• TNF is a cytokine produced mainly by activated macrophages, and is the
major extrinsic mediator of apoptosis.
• Most cells in the human body have two receptors for TNF: TNF-R1 and TNF-
R2.
• The binding of TNF to TNF-R1 has been shown to initiate the pathway that
leads to caspase activation via the intermediate membrane proteins TNF
receptor-associated death domain (TRADD) and Fas-associated death
domain (FADD).
• The link between TNF and apoptosis shows why an abnormal production of
TNF plays a fundamental role in several human diseases, especially
autoimmune diseases
Caspase
• Many pathways and signals lead to apoptosis, but there is only one
mechanism that actually
• causes the death of a cell, namely the activation of proteolytic
caspases
• Caspases (cysteine aspartic acid proteases) are a family of cysteine
proteases, first synthesised as inactive procaspases.
• Eleven caspases have so far been identified in humans, taking one of
two forms:
• the initiator (apical) caspases and the effector (executioner) caspases.
• Initiator caspases (e.g. CASP2, CASP8, CASP9 and CASP10) cleave
inactive pro-forms of effector caspases, thereby activating them.
• Effector caspases (e.g. CASP3, CASP6 and CASP7) in turn cleave other
protein substrates within the cell, to trigger the apoptotic process.
• The initiation of this cascade reaction is regulated by caspase
inhibitors.
• Caspases are regulated at a post-translational level, ensuring that
they can be rapidly activated.
P53
• p53 is a transcription factor which in humans is encoded by the TP53
gene, located on the short arm of chromosome 17 (17p13.1).
• It regulates the cell cycle and thus functions as a tumour suppressor
involved in preventing cancer; p53 has been described as ‘the
guardian of the genome’.
Thank you