ORGANELLES AND CELLULAR STRUCTURES

Cell theory:
1. Cells are the smallest unit of life
2. All cells arise from pre-existing cells by dividing into 2
3. All known living things are made of one or more cells.

Nucleus:

Diagnostic features:
● Enclosed by a nuclear envelope
- Nuclear envelope is a double membrane. The 2 membranes are separated by a
narrow space, known as perinuclear space.
- Outer membrane is continuous with the endoplasmic reticulum
- Nucleoplasm, a semi-fluid matrix, that fills the nucleus
- Perforated with nuclear pores that regulates entry and exit of substances into and
out of the nucleus
● Contains chromatin, coils of negatively charged DNA wrapped around positively charged
histone proteins
- Heterochromatin: electron dense, and appears as coarse granules under the electron
micrograph because it is more tightly packed
- Euchromatin: a less compact form of chromatin and is often under active
transcription
● Contains nucleolus, a prominent, dense, spherical structure in a non-dividing nucleus
- Contains large loops of DNA from a number of chromosomes. Loops contains genes
coding for a specific type of RNA known as ribosomal RNA

Functions:
Nucleus:
1. Stores hereditary material (DNA)
2. Controls gene expression. Site of transcription

Nucleolus:
1. Site of rRNA synthesis
 2. Assembly of proteins and rRNA, where proteins imported from the cytoplasm are
assembled with rRNA to form small and large ribosomal subunits.

Nuclear pores:
1. Regulates exchange of substances between the nucleus and cytoplasm, thus effectively
controlling processes on either side of the nuclear membrane.

Rough endoplasmic reticulum:

Diagnostic features:
1. Comprises continuous network of sheets of membrane
2. Outer surface is studded with ribosomes

Function:
1. Site of synthesis of proteins destined for secretion of incorporation into membranes
2. Facilitation of folding of polypeptide chain

Smooth endoplasmic reticulum:

Diagnostic features:
1. Comprises network of membranous tubules
2. Lacks ribosomes

Function:
1. Synthesis of lipids, including phospholipids, oils and steroids
2. Detoxification of drugs and poisons

Golgi Apparatus:

Diagnostic feature:
1. Consists of stack of flattened membrane-bound sacs called cisternae
2. Comprises a system of associated vesicles, Golgi vesicles
Functions:
1. Further modifies products of the ER, sorts and packages them into vesicles to be transported
into other parts of the cell, or to be released out of the cell
2. Synthesis of secretory polysaccharides
3. Synthesis of lysosomes

Polypeptides are synthesised at the ribosomes on the RER. These polypeptides enter the lumen of
RER and fold into proteins. The proteins are then modified and packaged into transport vesicles
which pinched off from RER. Transport vesicles then fuse with the cis face of the Golgi Apparatus
and release the proteins into the cisternae of the Golgi Apparatus. The proteins are further
modified, sorted and packaged into secretory vesicles at the Golgi Apparatus. The secretory vesicles
containing the modified protein are pinched off from the trans of Golgi apparatus. They then move
to fuse with the cell surface membrane. Secretory vesicles fuse with the cell surface membrane and
protein is released out of the cell via exocytosis.

Lysosomes:

Diagnostic features:
1. Spherical sac bound by a single membrane
2. Uniformly granular electron-dense appearance in electron micrographs
3. Constraints hydrolytic enzymes that digest macromolecules

Functions:
1. Digestion of material taken in from the environment by endocytosis
2. Autophagy: degradation of unwanted structures within the cell
3. Autolysis: self-destruction of a cell

Mitochondria:

Diagnostic features:
1. Wall comprises 2 membranes separated by an extremely narrow fluid-filled space called the
intermembrane space
- Outer membrane is smooth
- Inner membrane is highly convoluted with numerous infoldings called cristae
 2. Interior of mitochondria is an organic, semifluid matrix (mitochondrial matrix) with 70s
ribosome, circular DNA and various enzymes
3. The mitochondrial space is separated into 2 compartments: mitochondrial matrix and
intermembrane space

Function:
1. Main site of ATP production during aerobic respiration

Chloroplast:

Diagnostic features:
1. Enclosed by a double membrane known as chloroplast envelope, with a very narrow
intermembrane space separating the two membranes
- Inner membrane encloses a semi-fluid material known as stroma
- Stroma contains enzymes, circular DNA, 70s ribosomes, starch grains, sugars and
lipid granules
2. In the stroma is another membranous system in the form of flattened, interconnected
disc-like sacs called thylakoids
- The thylakoids are arranged in stacks called grana
- Stacks of grana are linked by intergranal lamellae
- Photosynthetic pigments and enzyme systems involved in photosynthesis are
embedded in the thylakoid membranes
3. The membranes of the chloroplast divide the chloroplast space into three compartments:
the intermembrane space, stroma and thylakoid space

Function:
1. Site of photosynthesis

Ribosomes:

Diagnostic features:
1. Complexes of ribosomal RNA and protein
2. 80s eukaryotic ribosomal consists of small (40s) and large (60s) subunits
3. Four different locations in the cell
 - Suspended as free ribosomes in cytosol
- Bound ribosomes attached to rough ER
- Mitochondrial matrix
- Chloroplast stroma

Function:
1. Site of polypeptide synthesis
2. Key role in translation of mRNA base sequence into specific amino acid sequence of a
polypeptide chain

Centrioles:

Diagnostic features:

1. Located in a region near the nucleus called the centrosome

2. Exist as a pair of rod-like structures positioned at right angles to each other
3. Transverse section reveals 9 triplets of microtubules arranged in a ring

Function:
1. Key role in nuclear division in animal cells by acting as microtubules organising centres

Plant cell wall:

Diagnostic features:
1. A rigid layer surrounding the cell, composed of insoluble cellulose fibres embedded in a
matrix of other polysaccharides and proteins
2. Middle lamella allows for adjacent cells walls to adhere to each other
3. Cell walls perforated with channels known as plasmodesmata

Functions:
1. Provides mechanical support to plant cells and maintain their shape
2. Allows development of turgor when water enters the plant cells by osmosis
3. Prevents excessive uptake of water
 CELLULAR TRANSPORT:

The fluid mosaic model of biological membrane comprises phospholipids and proteins that are
constantly moving freely within the layer in a lateral motion, making it fluid. Proteins are
interspersed and randomly embedded in a phospholipid bilayer giving it a mosaic appearance.

As temperature increases, membrane fluidity decreases.

Generally, the longer the fatty acid hydrocarbon chains, the higher the melting point of the
membrane. (membrane fluidity decreases) This is because a longer chain length increases the
surface area of contact, thus increasing the tendency of the hydrocarbon tails interacting with one
another via hydrophobic interactions.

Unsaturated fatty acid chains with cis carbon-carbon double bonds have kinks. These kinks hinder
the hydrocarbon chains from packing closely together, thus increasing membrane fluidity.

Saturated fatty acid chains are long and straight hydrocarbon chains. This facilitates close packing,
thus decreasing membrane fluidity.

Cholesterol:
● At higher temperatures, cholesterol limits fluidity. This is because cholesterol partly
immobilises adjacent phospholipid molecules, thus restraining their mobility. This
contributes to membrane stability at higher temperatures.

● At low temperatures, cholesterol interferes with the close packing of the phospholipids,
therefore enhancing membrane fluidity.

Functions of glycoproteins and glycolipids:

1. Serving as specific cell surface markers for cell-cell recognition processes
2. Cell-cell adhesion for purposes such as forming tissues
3. Serving as specific receptors sites for hormones and neurotransmitters
4. Keeping various other cells at a distance, preventing unwanted protein-protein interactions
Membrane function:
1. Acting as a boundary between intracellular and extracellular boundary
2. Selective barrier regulating passage of substances into and out of cells and organelles
3. Signal transduction
4. Cell-cell adhesion and recognition
5. Site of formation of multi-enzyme complexes
6. Compartmentalisation:
- Provides localised environmental conditions within the cell for specific metabolic
processes to take place simultaneously
- Enables cell to maintain high concentrations of certain enzymes and substrates in
specific compartments
- Prevents intermediates of one pathway from interfering with one another
- Isolates harmful substances from the rest of the cell

Diffusion: Small, non-polar and uncharged

Net movement of substances across the lipid
bilayer through transient gaps between
phospholipid molecules in the membrane,
down a concentration gradient, without
additional investment of ATP by the cell

Facilitated diffusion: Small, polar or charged

Net movement of substances from a region
from a higher concentration to a region of
lower concentration, through a selectively
permeable membrane, with the help of
transport proteins and without additional
investment of ATP by the cell

Osmosis: Small polar and uncharged

Net movement of water molecules from a
region of lower water potential to a region of
higher water potential, through a selectively
permeable membrane, without additional
 investment of ATP by the cell

Active transport: Small, charged or charged

Movement of particles from a region of lower
concentration gradient to a region of higher
concentration gradient, across a selectively
permeable membrane, with the additional
investment of ATP by the cell

Endocytosis: Macromolecules
Movement of large quantities of
macromolecules into the cell, via invagination
of the membrane, enclosing the
macromolecules, pinching off as a vesicle and
membrane reforms with the additional
investment of energy

Exocytosis: Macromolecules
Movement of large quantities of
macromolecules out of the cell. The vesicle
containing macromolecules move to the cell
surface membrane, membrane of the vesicle
fuses with a small portion of the cell surface
membrane, enabling macromolecules to be
released out of the cell via exocytosis, and
membrane reforms with the additional
investment of energy.

Phagocytosis:
1. A small portion of the cell surface membrane invaginates to enclose the solid particle
2. The particle is packaged within a membrane-enclosed sac that is large enough to be called a
vacuole
3. Phagocytic vacuole pinches off from the cell surface membrane
4. Cell surface membrane reforms
Receptor-mediated endocytosis:
1. When the specific substance, known as ligands, binds to the specific receptor sites of
receptor proteins, they trigger the invagination of the cell surface membrane
2. This results in the formation of coated vesicles that contain specific ligand molecules
3. After the ligands are taken up by the cells, the receptors are brought back to the cell surface
membrane by the same vesicles

CARBOHYDRATES:

a-glucose b-glucose

Structure: Can exist as both linear and ring forms

6 carbon sugar

Carbon atom 1 combines with oxygen on carbon atom 5 to give

pyranose ring

Hydroxyl group is below the Hydroxyl group is above the plane

plane of the ring of the ring

The ratio of hydrogen atoms to oxygen atoms is always 2:1

Every carbon atom except for C5 is attached to an -OH group and a

hydrogen atom

Property: Soluble in aqueous mediums

Has reducing powers

Condensation:
During condensation reaction, the hydroxyl group of carbon 1 of one glucose molecule and the
hydroxyl group of another glucose molecule’s carbon 4 react to form a glycosidic bond with the
elimination of one water molecule, catalysed by enzymes.
Elimination:
During hydrolysis, bonds between glucose monomers are broken by the addition of water; with a
hydrogen atom from the water molecule attaching to carbon 4 of one glucose monomer, and the
hydroxyl group from the water molecule attaching to carbon 1 of another glucose monomer.

- Chemical method: incubating with dilute acid at 100 degrees celsius

- Enzymatic method: incubating with an enzyme at room temperature
Starch and Glycogen:
1. Large molecules comprising several 100s/1000s of glucose residues → insoluble in water and
stores large amounts of energy
2. Glucose residues are joined by a-1,4/ a-1,6 glycosidic bonds → the bonds result in helical coil
so that structure is more compact and ideal for storage
3. Anomeric carbon of each glucose monomers is involved in glycosidic bond formation →
stable/ unreactive compound
4. Amylopectin is highly branched but for glycogen, it is even more highly branched with
shorter branch chains → branching allows many enzymes to act on amylopectin at one time,
therefore, many sites for hydrolysis. This allows rapid release of glucose residues

Cellulose:
1. Large molecule composed of several thousands of glucose monomers → insoluble in water
and stores energy
2. Alternate glucose monomers are inverted. Glucose residues are linked by b-1,4 glycosidic
bonds → the bonds result in long, straight chains
3. Hydroxyl groups project outwards from cellulose chain in all directions → allows
microfibrils to be arranged in large bundle to form macrofibrils → resulting in high tensile
strength

LIPIDS:

Condensation:
3 fatty acid molecules combine with 1 glycerol molecule. Each hydroxyl group in the glycerol
molecule reacts with the carboxyl group of a fatty acid. An ester bond is formed between the
glycerol and fatty acid, with the removal of a molecule of water - condensation reaction. Since
glycerol has 3 hydroxyl groups, 3 fatty acids attach themselves to the glycerol molecule and hence, 3
molecules of water are removed. Reaction is catalysed by enzymes.

Hydrolysis:
3 ester bonds are broken by 3 hydrolysis reactions with the addition of 3 water molecules. The H
atom of each water molecule attaches to glycerol and the OH group of each water molecule attaches
to a fatty acid. This reaction is catalysed by enzymes.

Triglycerides:

Triglyceride molecules are large and uncharged Can be stored in large amounts without having
- Insoluble in water any effect on the water potential of the cells

Large amounts of hydrogen atoms in their Storage of energy: they store more energy per
hydrocarbon chains which can be oxidised unit mass than any other respiratory substrates
during cellular respiration

Large amounts of hydrogen atoms in their Source of metabolic water

hydrocarbon chains which can be oxidised to
water

Less dense than water Confers buoyancy

Provide thermal insulation Prevents excessive heat loss, especially for

animals living in cold climates

Provide electrical insulation Component of myelin sheath, which allows

rapid transmission of nerve impulses

Forms natural buffer Absorbs shock, helps cushion the fragile,

internal organs from impact and physical
damage

Phospholipid:

The phosphate group of the phospholipid The hydrophobic region of the bilayer forms a
forms the hydrophilic “head” of the molecule barrier between the aqueous interior and
 whilst the hydrocarbon chains of the fatty acid exterior of the cell, whilst the hydrophilic
moieties form the hydrophobic “tails” of the regions of the bilayer make possible the
molecule. existence of a hydrophobic boundary.

In a phospholipid bilayer, the hydrophilic

“heads” of the phospholipid molecules make
contact with the aqueous environment whilst
the hydrophobic “tails” of the molecules are
shielded from the aqueous medium.

PROTEINS:

Condensation:
The carboxyl group of one amino acid and the amino group of another amino acid reacts to form a
peptide bond in a condensation reaction, with the elimination of a water molecule. The reaction is
catalysed by enzymes

Hydrolysis:
1 peptide bond is broken by 1 hydrolysis reaction with the addition of one water molecule. The H
atom of the water molecule attaches to one amino acid, while the hydroxyl group of the water
molecule attaches to another amino acid. The reaction is catalysed by enzymes.

Primary structure Specific sequence of amino acids Covalent peptide bonds

between successive amino acid
residues

Secondary structure A-helix: Hydrogen bonds

Maintained by hydrogen bonds
between CO and NH groups of
amino acid residues

B-pleated sheet:
 Maintained by hydrogen bonds
between adjacent regions of a
polypeptide chain that lie parallel to
each other, either running in the
same or opposite direction

Tertiary structure The polypeptide chain usually bends R group interactions:

and folds extensively, forming a 1. Disulfide bonds:
precise, compact, globular shape sulfhydryl groups are
oxidised to form a
Quaternary structure Many proteins consisting of 2 or
disulfide bond
more polypeptide chains
2. Ionic bonds: between
acidic and basic R
groups (oppositely
charged)
3. Hydrogen bonds:
between polar R
groups
4. Hydrophobic
interactions: between
nonpolar R groups

Haemoglobin:

The haemoglobin molecule is compact and Many haemoglobin molecules can be packed
globular in shape into a red blood cell for transport of oxygen

Each haemoglobin molecule consists of 4 Each haemoglobin molecule can carry up to 4

subunits: each capable of binding to a oxygen oxygen molecules, which greatly facilitates
molecule transport of oxygen

Haemoglobin is an allosteric protein that Maximises the amount of oxygen loaded and
exhibits cooperative binding of oxygen released at the lungs

The polypeptide chain of the subunit folds in Haemoglobin is soluble in red blood cells
 such a way that the hydrophobic amino acids
are buried on the interior while the hydrophilic
amino acid residues are on the exterior

Globin contains a deep hydrophobic cleft, ie. Provides a hydrophobic environment for the
haem-binding site binding of haem

Haem group contains a porphyrin ring and a Fe2+ can combine reversibly with oxygen, thus
Fe2+ ion enhancing the release of oxygen into
metabolically active tissues

Tropocollagen:

Tropocollagen molecules have a large Collagen is insoluble in water

molecular size. The polypeptide chains also
consist largely of glycine and proline which are
hydrophobic in nature.

Every third amino acid of each polypeptide Allows the three polypeptide chains to lie close
chain is a glycine together to form a tight coil

Each polypeptide chain contains many proline The bulky and inflexible proline and
and hydroxyproline residues hydroxyproline residues contribute to the
rigidity of the molecule

Within the tropocollagen molecule, extensive Results in high tensile strength for its structural
cross linking of hydrogen bonds between the role
NH group of glycine and CO group of
hydroxylysine holds the three polypeptide
chains together to form the triple helix.

Many tropocollagen molecules lie parallel to

each other in a staggered arrangement and
cross-linked via covalent bonds between the
carboxyl end of one molecule and the amino
 end of another. The tropocollagen bundled to
form fibrils which further assemble to form
collagen fibres.

ENZYMES:

Substrate alignment:
The enzyme molecule holds the different substrate molecules in an arrangement that forces close
together in the correct orientation. The proximity of the substrates within the enzyme-substrate
complex greatly increases the probability of a reaction occurring.

Bonds in substrate under physical stress:

Once a substrate binds to the active site of an enzyme, certain bonds within the substrate molecule
may be placed under physical stress. This increases the likelihood that the bonds will break.

Substrates acquired charged region:

When the R groups of amino acid residues at the active site are very close to part of the substrate,
the R groups can
1. Change the charge on the substrate, altering the distribution of electrons within the bonds
of the substrate
2. Cause other changes that will increase the reactivity of the substrate.
Enzyme concentration:

At low enzyme concentrations:

As enzyme concentration increases, the frequency of successful collisions between the enzyme and
substrate molecules increases. More enzyme-substrate complexes are formed per unit time, thus the
rate of reaction increases. Hence the rate of reaction is limited by enzyme concentration.

At high enzyme concentrations:

If substrate concentration was the limit factor, any further increase in enzyme concentration would
not result in an increase in rate of reaction.

Substrate concentration:
At low substrate concentrations:
As substrate concentration increases, the frequency of successful collisions between enzymes and
substrates would increase. More enzyme-substrate complexes are formed per unit time and rate of
reaction increases. Hence the rate of reaction is limited by enzyme concentration.

At high substrate concentrations:

The active sites of the enzyme molecules are saturated with substrate molecules. Any extra substrate
molecule has to wait until the enzyme-substrate complex has released the product before it can
enter the active site of the enzyme. The rate of reaction is now limited by enzyme concentration.

Temperature:

As temperature increases, the average kinetic energy of enzyme and substrate molecules increases.
There is an increase in frequency of successful collisions between enzyme and substrate molecules,
more enzyme-substrate complexes are formed per unit time and rate of reaction increases.

Above the optimum temperature, the atoms which make up the enzyme molecule vibrate so
vigorously that the hydrogen bonds and hydrophobic interactions between the R groups of the
amino acid residues begin to break. The enzyme is said to be denatured and the shape of its active
site is no longer complementary to that of the substrate.

pH:

At optimum pH, the intramolecular bonds maintaining the 3D confirmation of the enzyme
molecule are still intact. Majority of enzymes have the shape of the active site most ideal for binding
with substrate. Frequency of successful collisions between enzymes and substrate molecules is the
highest. Hence, the number of enzyme-substrate complexes formed per unit time is the highest and
rate of reaction is the fastest.

Above optimum pH, change in concentration of H+ in the environment alters the charges in the
acidic and basic R groups of the amino acid residues. This disrupts ionic bonds and hydrogen bonds
between R groups of amino acid residues. The shape of the active site is no longer complementary
to that of the substrate. Enzymes are denatured. Hence, less/no enzyme-substrate complexes are
formed per unit time and the rate of reaction decreases.

Competitive inhibition:

● Similar shape as the substrate

● Compete for the same active site
● Reduces the rate of reaction by lowering the proportion of enzyme molecules bound to the
substrate
● Effect of competitive inhibition can be overcome by increasing substrate concentration
- Increased substrate concentration increases the frequency of successful collisions
between enzyme and substrate molecules, rather than enzyme and inhibitor
- At very high substrate concentrations, the rate of reaction can reach the same as
maximum value as that in the absence of inhibitors

Non-competitive inhibition:

● No structural similarity to the substrate. It binds at a region other than the active site
● Renders a proportion of enzyme molecules out of action by decreasing the number of
enzyme-substrate complexes formed per unit time
● Not reach maximum value as that in the absence of inhibitors
● Cannot be overcome by increasing substrate concentration

DNA REPLICATION:

Each nucleotide is composed of a nitrogenous base, pentose sugar and phosphate group.

Pentose:
- A five carbon sugar that occurs as a ring form
- Ribose in RNA and deoxyribose in DNA

Nitrogenous base:
- Heterocyclic ring of carbon and nitrogen atoms
- Purines: double ring (adenine, guanine)
 - Pyrimidine: single ring (cytosine and thymine)

Physical structure of DNA:

1. Thin, long molecule with a diameter of 2nm
2. Double-stranded
3. Coiled in the form of a helix, the helix makes a complete turn every 3.4nm
4. 10 bases to each complete turn
5. Purines = pyrimidines >> C+T = A+G
6. A+T : C+G >> 1:1
7. A:T >> 1:1 and C:G >> 1:1

Main features of the double helix:

1. DNA consists of 2 polynucleotide strands
2. Each polynucleotide strand forms a right-handed helical spiral and the two strands coil
around each other to form a double helix
3. The diameter of the double helix is 2nm
4. The two polynucleotide strands run in opposite directions, ie. They are antiparallel. One
strand is oriented in the 5’ to 3’ direction while the other is oriented in the 3’ to 5’ direction.
5. Each strand has a sugar phosphate backbone:
- Phosphate groups project outside the double helix
- Nitrogenous bases that orientate inwards the central axis
6. Specific complementary base pairing occurs between A&T and C&G
7. A-T base pair is held by 2 hydrogen bonds, while C-G base pair is held by 3 hydrogen bonds
8. A:T is 1:1 and C:G is 1:1
9. Along the central axis of the DNA molecule, the base pairs are stacked 0.34nm apart. One
complete turn of the double helix has 10 base pairs and spans a distance of 3.4nm.
10. Because of the way the bases bond with each other, the two sugar phosphate backbones of
the double helix are not equally spaced along the helical axis.
- This results in grooves of unequal sizes between the backbones called major and
minor grooves being formed

Stability of the DNA molecule:

1. Phosphoester bonds between nucleotides are strong covalent bonds
2. A large number of hydrogen bonds between base pairs
 3. Hydrophobic interactions between the stacked base pairs
4. The exposure to outside influences of only the sugar phosphate backbone
5. Involvement of nitrogenous bases in hydrogen bonds
6. The DNA double helix is tightly wound around an equal mass of histones to form a
repeating array of nucleosomes

Semi-conservative DNA replication: this involves the separation of parental DNA strands, whereby
each strand acts as a template for the synthesis of a new daughter strand. The daughter molecule
therefore comprises one parental strand and one new daughter strand.

DNA Replication:

1. Replication of a DNA molecule begins at specific sites called origins of replication

2. The two polynucleotide strands separate and form a replication bubble. At the end of each
replication bubble is a replication fork.
3. Replication of DNA proceeds in both directions until the entire molecule is copied.

1. Helicases catalyses the breakage of hydrogen bonds, thus separating the two parental DNA
strands
2. Single-stranded binding proteins bind tightly to the single-stranded regions of DNA to help
maintain the stability of the replication fork
3. Topoisomerase creates a transient break by nicking a strand of DNA. This helps to unwind
the double helix ahead of the replication fork for initiation of replication

1. A portion of each parental DNA strand serves as a template for making the RNA primer
with complementary base sequence.
2. An enzyme called primase catalyses the synthesis of RNA primer in the 5’ to 3’ direction
3. DNA polymerase (III) can now elongate the strand by adding the next dNTP to the free 3’
hydroxyl group of the primer.
4. DNA polymerase (I) will later replace RNA nucleotides of the primers with DNA
nucleotides

1. The separated parental DNA strands form the template along which deoxyribonucleoside
triphosphate (dNTP) align themselves by complementary base pairing
 2. DNA polymerase (III) catalyses the formation of a phosphoester bond between the 3’
hydroxyl group of the last nucleotide in the growing strand and the 5’ phosphate group of
the incoming dNTP
3. DNA polymerases (III) thus catalyses the polymerisation of the new DNA strand in the 5’ to
3’ direction
4. Each growing new DNA strand is antiparallel to its parental template strand
5. The leading strand is synthesised continuously as a single polymer along the template strand
6. The lagging strand is synthesis discontinuously as a series of short fragments called Okazaki
fragments along the template strand
7. DNA ligase catalyses the formation of a phosphoester bond between the 3’ end of each new
Okazaki fragment and the 5’ end of the growing strand to form a continuous strand
8. Each daughter DNA molecule now consists of a parental strand and a newly synthesised
strand

End replication problem:

The end replication problem exists due to the specificity of DNA polymerase. Both the 3’ hydroxyl
group and 5’ phosphate group are required to fit into the enzyme active site for the reaction to
occur. Without a 3’ hydroxyl group at the 5’ ends of the newly synthesized strand, the addition of
new DNA nucleotides is not possible, resulting in a gap.

This shortening of DNA strands after every replication limits the number of times a cell can divide
before apoptosis is triggered.

Telomeres found at the ends of DNA molecules are used to buffer the loss of important genetic
information as a result of this end replication loss.

TRANSCRIPTION:

1. RNA polymerase recognises and attaches to the promoter of the gene on DNA
2. RNA polymerase breaks the hydrogen bonds between complementary base-pairs of the
DNA double helix
3. The DNA double helix unwinds and the two DNA strands separates
4. One of the DNA strands acts as a template for the formation of mRNA
 1. Free ribonucleoside triphosphates are aligned along the DNA template strand
2. According the complementary base-pairing rule
3. RNA polymerase catalyses the linkage of ribonucleotides via phosphoester bond
4. RNA polymerase catalyses elongation of the RNA in the 5’ to 3’ direction by adding
ribonucleotides to the free 3’OH end of the growing RNA molecule
5. The separated DNA strands rewind into the double helix behind the RNA polymerase

1. Transcription proceeds until after the RNA polymerase transcribes a termination sequence
in the DNA
2. RNA polymerase detaches from the DNA molecule. mRNA is released

Post-transcriptional modifications:

The 5’ end is immediately capped off with a modified form of guanine nucleotide. This 5’ cap has at
least 2 important functions:
a. Helps protect the mRNA from degradation by hydrolytic enzymes
b. After the mRNA reaches the cytoplasm, the 5’ cap functions as part of an “attach here” sign
for ribosome

At the 3’ end, an enzyme makes a poly(A) tail consisting of some 50 to 250 adenine nucleotides
a. Serves to inhibit degradation and probably helps ribosomes attach to it
b. Facilitate export of mRNA from the nucleus
Spliceosome cuts out the introns from the pre-mRNA and joins all the exons together into a
continuous coding strand via splicing.

Genetic code:
T: genetic code occurs in triplet of bases
U: genetic code is universal ie. the same codons apply for the same amino acids
N: genetic code is non-overlapping: the mRNA sequence is read continuously without skipping any
nucleotides
D: genetic code is degenerate, >1 codon may code for the same amino acid
Start codon: AUG
Stop codon: UGA, UAA, UAG

Structure-function of tRNA

Anticodon at one end Specifies the identity of the amino acid at the 3’
end

Specific sequence of bases in amino acid Allows complementary base pairing with
corresponding codon mRNA

3’CCA end of tRNA Attachment site of specific amino acid

tRNA adopts a compact L shape Anticodon lies at one end and the point of
attachment of the amino acid lies on the end
end: to reduce steric hindrance during
translation

tRNA has 3 loops 1. Forming complementary base pairing

with rRNA of ribosome
2. Binding with amino-acyl tRNA
synthetase during amino acid activation
3. Binding to specific mRNA codon via
complementary base pairing

Ribosome:
1. Small subunit has a binding site for mRNA
2. Large subunit has 3 binding sites for tRNA
a. peptidyl-tRNA site holds the tRNA carrying the polypeptide chain
b. aminoacyl-tRNA site holds the tRNA carrying the next amino acid to be added to
the polypeptide chain
c. Exit site allows the discharged tRNA to leave the ribosome

TRANSLATION:
 1. An amino acid is attached to each tRNA molecule at its 3’ CCA end forming
aminoacyl-tRNA complex
2. tRNA molecules bind to their specific amino acid as determined by their anticodon
3. This reaction is catalysed by the enzyme aminoacyl-tRNA synthetase

1. mRNA molecule binds to a small ribosomal subunit

2. The anticodon of the initiator tRNA binds to the start codon of mRNA
3. The large ribosomal subunit then binds to the small ribosomal subunit forming the
initiation complex

1. Anticodon of incoming tRNA molecule carrying its amino acid pairs with the mRNA codon
in the aminoacyl-tRNA site of the ribosome
2. Peptidyl transferase catalyses the peptide bond formation between the amino acid carried by
the tRNA in the A site with the amino acid bound to tRNA in peptidyl-tRNA site
3. The ribosome translocates the tRNA in the A site, with its attached polypeptide, to the P
site, taking the mRNA with it
4. The mRNA is moved through the ribosome from the 5’ to 3’ direction on the mRNA
5. Elongation cycle is repeated until the ribosome reaches a stop codon

1. Release factor binds directly to the stop codon in the A site

2. Release factor causes the addition of a water molecule
3. This reaction hydrolyses the completed polypeptide from the tRNA and free the polypeptide
from the ribosome
4. Translational complex is disassembled
5. Polypeptide chain folds into its unique 3-dimensional shape

EUKARYOTES:

First level of condensation:

1. DNA is packed into nucleosome to produce the 10-nm interphase chromatin fibre =
nucleosome fibre
2. A nucleosome consists of DNA wound twice around a protein core composed of an octamer
of 4 histone molecules
 3. In the cell cycle, the histones leave the DNA only briefly during DNA replication and gene
expression processes that require access to the DNA by the cell’s molecular machinery

Second level of condensation:

1. DNA is further folded or coiled to produce the 30-nm chromatin fibre = solenoid
2. Histone H1 and linker DNA is involved in this coiling of the 10-nm nucleosome fibre to
produce the 30-nm chromatin fibre

Third level of condensation:

1. Non-histone chromosomal proteins form a scaffold that is involved in condensing the
30-nm chromatin fibre into loops called looped domains, forming 300-nm fibre
2. In mitotic and meiotic chromosomes, the looped domains themselves coil and fold, further
compacting all the chromatin to produce the characteristic metaphase chromosome. The
width of one chromatid is 700nm
3. Particular genes always end up located at the same places in mitotic and meiotic
chromosomes, indicating that the packing steps are highly specific and precise.

Coding sequences:
Exons

Non-coding sequences:
1. Introns
2. 5’ and 3’ untranslated regions (UTR)
3. Promoters
4. Enhancers and silencers
5. Replication origin and termination
6. Promoters and telomeres

● Composed of repeated sequences:

- Tandemly repetitive DNA: centromeres and telomeres
>> Transcriptionally inactive and play a structural role in the chromosome

- Moderately repetitive DNA: promoters, enhancers and origins of replication

>> scattered about the genome
Centromere:
Structure:
The centromere is a repetitive DNA sequence. It is an A-T rich region ranging in size from 102 to
106 base pairs.

Function:
The centromere binds several proteins with high affinity. This complex, called the kinetochore,
provides the anchor for the spindle fibres. The centromere is thus an essential structure for
chromosome segregation during cell division.

Telomere:
Structure:
Telomeric sequences are repetitive DNA sequences. They are typically short, repeated T-G rich
sequences.

Function:
1. To prevent fusion of the ends of the chromosomes
2. To prevent exonucleases from degrading the ends of the DNA molecules
3. To prevent the loss of genetic information by acting as a disposable buffer
4. To facilitate replication of the ends of the linear DNA
5. To prevent cell cycle arrest

PROKARYOTIC CHROMOSOME EUKARYOTIC CHROMOSOME

Circular chromosomal DNA Linear DNA

Few millions base pairs in length Tens of millions to hundreds of millions in

length

2 sets of chromosomes Single type of chromosome but may be present

in multiple copies

Several thousand different genes Few hundred and several thousand different
 Interspersed throughout the chromosome genes
Interspersed throughout the chromosome

One origin of replication Many origins of replication

Short repetitive sequences but may but - Contains a centromere that forms a
interspersed recognition site for kinetochore
- Telomeres contain specialised
sequences
- Repetitive sequences are commonly
found near centromeric and telomeric
regions but may be interspersed

CONTROL:

Histone acetylation:
● In acetylation, acetyl groups are attached to lysines in histone tails. The enzyme catalysing
the transfer is histone acetyltransferase.
● When lysines are acetylated, their positive charges are neutralised. As such histone tails no
longer interact with the neighbouring nucleosomes.
● Electrostatic interaction between neighbouring nucleosomes promotes folding of chromatin
into a more compact structure.
● Histone acetylation does not only promote transcription initiation by remodelling the
chromatin structure, the enzyme HAT also binds to and aids in the recruitment of the
transcription machinery

Histone deacetylation:
● Histone deacetylases >> remove acetyl groups from lysines in histone tails >> form a
condensed structure >> no transcription

Histone Methylation:
● The addition of methyl groups to histone tails promotes condensation of chromatin >> no
transcription
DNA methylation:
● Cytosine residues can be methylated to produce 5-methylcytosine. These methylated
cytosine residues are located in GC-rich sequences, which are often near or in the promoter
region of a gene
● Comparison of the same genes in different tissues shows that genes are usually more heavily
methylated in the cells in which they are not expressed.
● Moreover, proteins that bind to methylated DNA can recruit deacetylation enzymes. DNA
methylation and histone deacetylation work together to repress transcription

Transcription occurs:
1. Histone acetylation
2. Histone demethylation
3. DNA demethylation

Transcription suppressed:
1. Histone deacetylation
2. Histone methylation
3. DNA methylation

Two types of control elements:

1. Promoters which comprises the core promoter and proximal control elements
2. Distal control elements which comprises enhancers and silencers

Core promoter contains the transcription start site and TATA box:
1. TATA box is an A-T rich consensus sequence found upstream of transcription start site
2. TATA box serves as the binding site of general transcription factors to facilitate binding of
RNA polymerase II to the promoter, for transcription to occur
3. Core promoter determines the basal level of expression

Proximal control elements located upstream from the core promoter:

1. CAAT boxes and GC rich sequences
2. Determine how frequent transcription occurs
Distal control elements contain enhancers and silencers:
1. Enhancer has a binding site for specific transcription factors called activators
Attachment of activator to enhancer increases the rate of transcription of the gene
2. Silencer has a binding site for specific transcription factors called repressors
Attachment of silencer to repressor inhibits or decreases the rate of transcription

Post-transcriptional modifications:
1. 5’ capping: A modified guanosine triphosphate residue is added to the 5’ nucleotide of the
pre-mRNA to create a “capped” RNA
2. 3’ polyadenylation: The 3’ end of the pre-mRNA is cleaved and 50-250 adenine nucleotides
are added to the new 3’ end to create the poly(A)taill

Functions:
A. Facilitate export of mRNA from the nucleus
B. Protect the mRNA from degradation by hydrolytic enzymes
C. Help ribosomes attach to the 5’ end of mRNA once the mRNA reaches the nucleus

RNA Splicing:
➔ Non-coding sequences are known as introns and coding sequences are known as exons.
➔ The introns are cut out and the exons are joined together to form a continuous coding
sequence via mRNA splicing.

Significance of splicing:
The biological advantage lies in alternative splicing whereby a pre-mRNA can be spliced in more
than one way. Hence, alternative splicing produces two or more polypeptides with differences in
their amino acid sequences, leading to possible variation in their functions.
Factors affecting mRNA stability:

Length of poly(A)tail:
1. Longer poly(A)tail >> more stable, longer time to shorten the poly(A)tail by cellular
exonucleases >> increase stability of mRNA, translation occurs repeatedly
2. Shorter poly(A)tail >> less stable, less time to shorten the poly(A) tail >>less translation, less
proteins synthesised
Destabilising sequences:
mRNAs contain sequences that act as destabilising elements >> most commonly found within the 3’
untranslated region >> consensus rich AUUUA is recognised by cellular proteins that bind to the
ARE >> influencing whether mRNA is rapidly degraded

Biochemical modifications:
1. Cleavage of initial insulin polypeptide forms the active hormones
2. Chemical modifications such as phosphorylation and glycosylation
3. Cell-surface proteins and secretory proteins must be transported to target destinations
inside and outside the cells to function
4. Regulation may occur at any of the steps involved in modifying or transporting a protein

Polymerase chain reaction:

Denaturation:
1. An excess or primer is mixed with DNA fragment to be amplified
2. The thermal cycler heats the mixture of primer and DNA fragment to 94 degrees celsius
3. At this temperature, hydrogen bonds holding the double-stranded DNA fragment break and
double-stranded DNA dissociates into single strands

Annealing of primers:
1. The solution is allowed to cool to 65 degrees celsius
2. As it cools, the single strands of DNA tend to reassociate into double strands
3. Because of the large excess of both the forward and reverse primers, primers base pair with
complementary sequences in the single-stranded DNA

Primer extension:
1. The thermal cycler then raises the temperature to 72 degrees celsius, the temperature at
which the Taq polymerase functions best
2. Using the primer, the heat stable polymerase copies the rest of the fragment as if it were
replicating DNA
 3. The primer is then lengthened into a complementary copy of the entire single-stranded
fragment
4. Because both DNA strands are replicated, there are now two copies of the original fragment
Gel electrophoresis:

1. The gel acts as molecular sieve to separate nucleic acids or proteins on the basis of size,
electrical charge and other physical properties
2. A gel material consisting of agarose is poured as a thin slab on a glass
3. After the gel has solidified, the researcher loads samples containing a mixture of
macromolecules of different sizes in wells formed at one end of the gel
4. The gel is bathed in an aqueous solution and electrodes are attached to both ends and an
electrical current is applied
5. The DNA molecules, which are negatively charged due to their phosphate groups, migrate
towards the positive electrode
6. The distance a DNA molecule travels is inversely proportional to its length. Larger
molecules travel more slowly through the pores in the gel, thus the smallest DNA fragments
travel the longest distance
7. The gel slows down the large molecules more than the smaller molecules because the large
molecules have more difficulty in moving through the pores in the gel
8. When the current is turned off, the DNA molecules in each sample are arrayed in bands
along a “lane” according to their size. The shortest molecules having travelled the furthest,
are in bands at the bottom of the gel.
9. These bands of separated molecules can be visualised with stains
10. The actual size of the DNA fragments is determined by comparison to migration distances
of known marker fragments

Southern blot and nucleic acid hybridisation:

● Identify within the smear the size of the particular fragment containing the gene of interest

1. Genomic DNA samples are isolated from 3 individuals

2. A restriction enzyme is added to the 3 samples of DNA to produce restriction fragments

1. The restriction fragments from each sample are then separated by electrophoresis
2. Each sample forms a characteristic pattern of bands
 1. These fragments will then be transferred by capillary action from the gel to a sheet of
nitrocellulose membrane
2. Capillary action pulls an alkaline solution upward through the gel and through a sheet of
nitrocellulose membrane laid on top of it, transferring the DNA to the membrane and
denaturing it
3. The single strands of DNA stick to the membrane, positioned in bands exactly as on the gel

1. The blot is exposed to a solution containing radioactively labelled probe

2. The probe is single-stranded DNA complementary to the DNA sequence of interest, and it
attaches by base-pairing to restriction fragments of complementary sequence.

1. A sheet of photographic film is laid over the blot

2. The radioactivity in the bound probe exposes the film to form an image, corresponding to
DNA bands containing the DNA that base-paired with the probe

DNA MUTATIONS:

DNA Mutations:
1. Point mutations: involves chemical changes in 1 base-pair of DNA
● Base-pair substitution: replacement of 1 base-pair
● Base-pair addition: insertion of 1 base-pair
● Base-pair deletion: removal of 1 base-pair
2. Chromosomal aberration
● Numerical aberration: changes in number of chromosomes
● Structural aberration: changes to structure of chromosomes

● Base-pair substitution:
○ Silent mutations: no effect on the amino acid sequence
○ Missense mutations: little effect on/ major change to final protein
○ Nonsense mutations: production of a stop codon

● Base-pair addition and deletion

 ○ Frameshift mutation: missense and nonsense mutations (must occur in multitudes of
3)

Silent mutation:
● No effect on the amino acid sequence >> degenerate nature of genetic code >> mutation is
not observed

1. Base-pair substitution
2. Change in DNA sequence
3. Change in corresponding mRNA codon but still codes for the same amino acid
4. No change in amino acid sequence of polypeptide chain
5. No change to properties of R group
6. No change to structure, properties and function of protein
7. Polypeptide chain folds to produce the same 3D conformation

Missense mutation:
● Two different outcomes: little effect on final protein or major change to the final protein

Little effect on protein:

1. Base-pair substitution
2. Change in DNA sequence
3. Change in corresponding mRNA codon, code for amino acid with similar properties
4. Change in amino acid sequence of polypeptide chain
5. No change in structure, property and function of protein

Major effect on protein:

1. Base-pair substitution
2. Change in DNA sequence
3. Change in corresponding mRNA codon, code for amino acid with different properties
4. Change in amino acid sequence of polypeptide chain
5. Change in structure, property and function of protein: less active/ non-functional protein

Nonsense mutation:
 ● Produces a stop codon >> premature termination of translation >> truncated polypeptide

1. Base-pair substitution
2. Change in DNA sequence
3. Change in corresponding mRNA codon, resulting in STOP codon
4. Premature termination of translation
5. Truncated polypeptide
6. Non-functional or no protein

Frameshift mutation:
● If base-pair addition and substitution does not occur in multitudes of 3 >> frameshift
mutation >> missense and nonsense mutation

Missense mutation:
1. Base-pair deletion
2. Change in DNA sequence
3. Shift in reading frame
4. Change in sequence of mRNA codons
5. Significant change in amino acid sequence of polypeptide chain
6. Change in structure, property and function of protein

Nonsense mutation:
1. Base-pair addition
2. Change in DNA sequence
3. Shift in reading frame
4. Results in STOP codon on mRNA
5. Premature termination of translation
6. Truncated polypeptide chain
7. Non-functional or no protein

Sickle cell anemia:

● In a normal adult, the normal haemoglobin A (HbA) has a quaternary structure consisting of
two a-globin and two b-globin chains
● In individuals with sickle cell anemia, HbS is produced instead
 ● Sickle cell anemia is caused by a single BASE-PAIR SUBSTITUTION on a gene on
chromosome 11 that codes for b-globin chain of the haemoglobin molecule
● The mutation results in thymine being replaced by adenine at the 17th nucleotide of the
original gene sequence
● In the mRNA produced, the 6th codon is changed from “GAA” to “GUA”
● As a result, a neutral valine residue replaces the negatively charged glutamic acid residue
➔ Valine is hydrophobic while glutamic acid is hydrophilic
➔ This decreases the solubility of deoxygenated HbS in the red blood cell
● Since HbS is less soluble than HbA, it is less efficient at binding to and transporting oxygen
● The effects of HbS occur at low oxygen concentrations
1. A sticky patch on the haemoglobin is exposed
2. Adjacent HbS molecules associate with each other via sticky patches
3. The HbS molecules polymerise and precipitate out of the solution to form crystalline
structures
4. This causes the red blood cells to change from a circular biconcave shape to a sickle
shape

Chromosomal aberration:
a. Deletion: loss of gene >> shorter chromosome formed
b. Duplication: addition of genes >> section of chromosome replicates
c. Inversion: a chromosome breaks at 2 locations and the middle portions inverts 180o before
rejoining
d. Translocation: a section of chromosome breaks off and becomes attached to another
chromosome

Down syndrome: an extra copy of chromosome 21 is found within an individual’s cells

● This can result from nondisjunction of the homologous chromosome 21, which fails to
separate to opposite poles of the cell during anaphase 1, or anaphase II of meiosis. This
results in the gamete carrying two copies of chromosome 21.
● Subsequent fusion of this gamete with a normal gamete results in a zygote with 3 copies of
chromosome 21
 CELL CYCLE:

Mitosis:

Prophase:

Behaviour of chromosomes:
- Chromatin fibres become more tightly coiled, condensing into discrete chromosomes
- Each duplicated chromosome now appears as 2 identical sister chromatids, joined together
at the centromere

Nucleus:
- Nucleoli disappears as their DNA passes to certain chromosomes
- During late prophase, the nuclear envelope breaks down into small vesicles which disperse

Spindle fibres:
- Outside the nucleus, duplicated pair of centrioles move towards opposite poles of the cell
- Short microtubules may be seen radiating from the centrioles
- Microtubules extending from each centrosome then invade the nuclear area. Chromosomes
attach to kinetochore microtubules via their kinetochores, and thus undergo active
movement.

Metaphase:

Behaviour of chromosomes:
- Chromosomes are aligned at the metaphase plate of the spindle
- Kinetochore microtubules attach sister chromatids to opposite poles of the spindle

Nucleus:
- Nucleus absent
Spindle fibre:
- Centrosomes are now at opposite poles of the cell

Anaphase:

Behaviour of chromosomes:
- The centromere of each chromosome separates into 2, causing sister chromatids of each
chromosome to separate
- Each chromatid becomes a daughter chromosome

Nucleus:
- Nucleus absent

Spindle fibres:
- Kinetochore microtubules shorten, pulling the daughter chromosomes to opposite poles
centromeres first
- Interpolar microtubules lengthen, causing spindle fibres to move apart and the cell to
elongate

Telophase:

Behaviour of chromosomes:
- The two sets of daughter chromosomes arrive at the opposite poles of the spindle
- Chromosomes decondense into chromatins

Nucleus:
- A new nuclear envelope reassembles around the chromosomes at each pole and the nucleoli
reappear

Spindle fibres:
- Microtubules dis-assemble
Meiosis:

Significance:
1. Genetic stability
2. Growth and development of zygote
3. Cell replacement: supplies new cells required to replace damaged or worn-out cells
4. Regeneration
5. Asexual reproduction

Meiosis I:

Prophase I:

Behaviour of chromosome:
- Chromatin becomes more tightly coiled, condensing into discrete chromosomes
- Homologous chromosomes pair up to form a bivalent via synapsis
- The bivalents shorten and thicken further, partly by coiling
- The homologous chromosomes appear to repel each other and partially separate
- A process known as crossing over occurs between non-sister chromatids of homologous
chromosomes >> new combinations of alleles
- The bivalents assume particular shapes

Nucleus:
- The nucleoli disappear and nuclear envelope disintegrates

Spindle fibres:
- Outside the nucleus, the mitotic spindle assembles between the two centrosomes, which
moves towards opposite poles of the cell
Metaphase I:

Behaviour of chromosome:
- Homologous pairs of chromosomes are aligned on the metaphase plate of the spindle, with
one chromosome in each bivalent facing each pole
- Independent assortment of homologous chromosomes occurs >> orientation of each
bivalent along the metaphase plate is random and independent

Nucleus:
- Nucleolus and nuclear envelope absent

Spindle fibres:
- Centromeres of chromosomes are attached to kinetochore microtubules via kinetochore
proteins

Anaphase I:

Behaviour of chromosomes:
- Homologous chromosomes move towards opposite poles of the spindle
- This separates the chromosomes into 2 haploid sets

Nucleus:
- Nucleolus and nuclear envelope absent

Spindle fibres:
- Kinetochore microtubules shorten, pulling homologous chromosomes, centromeres leading
towards opposite poles of the spindle
- Interpolar microtubules lengthen, causing spindle fibres to move apart and the cell to
elongate

Telophase I:

Behaviour of chromosomes:
 - Homologous chromosomes reach opposite poles of the cell, thus marking the end of meiosis
I
- Chromosomes decondense to form chromatin

Nucleus:
- Nucleolus reappears and nuclear envelope reforms
Spindle fibres:
- Spindle fibres disassemble

Meiosis II:

Prophase II:

Behaviour of chromosomes:
- Chromatin becomes more tightly coiled, condensing into discrete chromosomes

Nucleus:
- Nucleoli disappear and nuclear envelope disintegrates

Spindle fibre:
- Centrosomes move to opposite poles of the cells
- New microtubules appear and become attached to kinetochore region of the chromosomes
- The new spindle microtubules are orientated at right angles to the original spindle
microtubules of meiosis I

Metaphase II:

Behaviour of chromosomes:
- Chromosomes, each with its two sister chromatids, are aligned individually along the
metaphase plate

Nucleus:
- Nucleolus and nuclear envelope absent
Spindle fibres:
- Kinetochore microtubules attach each of the two sister chromatids of a chromosome to
opposite poles of the spindle

Anaphase II:

Behaviour of chromosomes:
- Centrosomes separate, sister chromatids separate at the centromeric region. Each sister
chromatid is now known was a daughter chromosome

Nucleus:
- Nucleolus and nuclear envelope absent

Spindle fibres:
- Kinetochore microtubules shorten, pulling the daughter chromosomes to opposite poles,
centromeres leading
- Interpolar microtubules lengthen, causing spindle fibres to move apart and the cell to
elongate

Telophase II:

Behaviour of chromosomes:
- Daughter of chromosomes reach opposite poles of the cell
- Chromosomes decondense to chromatin

Nucleus:
- Nuclear envelope reform around each nucleus and nucleoli reappears, completing the
formation of four daughter nuclei

Spindle fibres:
- Spindle fibres disassemble
Proto-oncogenes: codes for proteins that stimulate normal cell-division
>> Gain-in-function mutation
Oncogenes: excess protein [hyperactive protein degradation resistant protein]

Tumor suppressor gene: codes for proteins that inhibit normal cell-division
>> Loss-in-function mutation

→ loss of cell-cycle control >> uncontrolled cell division