BASIC CONCEPTS: CHROMOSOMES, CELL DIVISION,

GENE STRUCTURE, TRANSCRIPTION AND

TRANSLATION, STRUCTURE OF THE HUMAN GENOME

Presenters: SIFA HYERA & NADIA AHMED

Facilitator: Dr Likindikoki
 CHROMOSOMES
• In the nucleus of each cell, the DNA molecule is
packaged into thread-like structures called
chromosomes
• Each chromosome is made up of DNA tightly
coiled many times around proteins called
histones that support its structure.
• Chromosomes are not visible in the cell’s
nucleus—not even under a microscope—when
the cell is not dividing
 Chromosomes…
• However, the DNA that makes up chromosomes
becomes more tightly packed during cell division
and is then visible under a microscope.
• Each chromosome has a constriction point called
the centromere, which divides the chromosome
into two sections, or “arms.” The short arm of the
chromosome is labeled the “p arm.” The long
arm of the chromosome is labeled the “q arm.
Chromosomes…
 DNA
• DNA, or deoxyribonucleic acid, is the
hereditary material in humans and almost all
other organisms. 
• Most DNA is located in the cell nucleus (where
it is called nuclear DNA), but a small amount
of DNA can also be found in the mitochondria
(where it is called mitochondrial DNA or
mtDNA).
 DNA…
• The information in DNA is stored as a code made
up of four chemical bases: adenine (A), guanine
(G), cytosine (C), and thymine (T)
• DNA bases pair up with each other, A with T and
C with G, to form units called base pairs
• Each base is also attached to a sugar molecule
and a phosphate molecule
• Together, a base, sugar, and phosphate are called
a nucleotide
 DNA…
• Nucleotides are arranged in two long strands
that form a spiral called a double helix.
• The structure of the double helix is somewhat
like a ladder, with the base pairs forming the
ladder’s rungs and the sugar and phosphate
molecules forming the vertical sidepieces of
the ladder.
DNA Structure
 Mitochondrial DNA
• Although most DNA is packaged in
chromosomes within the nucleus,
mitochondria also have a small amount of
their own DNA
• Mitochondrial DNA contains 37 genes, all of
which are essential for normal mitochondrial
function. 
 Mitochondrial DNA…
• 13 of these genes provide instructions for
making enzymes involved in oxidative
phosphorylation.
• The remaining genes provide instructions for
making transfer RNAs (tRNAs) and ribosomal
RNAs (rRNAs)
 Gene
• A gene is the basic structural and functional
unit of heredity
• Genes, which are made up of DNA, act as
instructions to make up a proteins
•  In humans, genes vary in size from a few
hundred DNA bases to more than 2 million
bases.
Gene…
 CELL DIVISION
• Cell division is the process by which a
parent cell divides into two or more daughter cells
• occurs as part of a larger cell cycle
• In eukaryotes, there are two distinct types of cell
division:
– a vegetative division, whereby each daughter cell is
genetically identical to the parent cell (mitosis),
– a reproductive cell division, whereby the number
of chromosomes in the daughter cells is reduced by half
to produce haploid gametes (meiosis).
 Cell division…
• Prokaryotes undergo a vegetative cell
division known as binary fission, where their
genetic material is segregated equally into two
daughter cells
• All cell divisions, regardless of organism, are
preceded by a single round of DNA replication.
Cell division
•  Mitotic cell division enables sexually
reproducing organisms to develop from the one-
celled zygote, which itself was produced by
meiotic cell division from gametes.
• After growth, cell division by mitosis allows for
continual construction and repair of the organism
• The primary concern of mitotic cell division is the
maintenance of the original cell's genome.
• Before division can occur, the genomic
information that is stored in chromosomes
must be replicated, and the duplicated
genome must be separated cleanly between
cells.
• A great deal of cellular infrastructure is
involved in keeping genomic information
consistent between generations.
MITOSIS
 Mitosis..
• mitosis is a part of the cell cycle when
replicated chromosomes are separated into
two new nuclei
Cell cycle
 Cell Cycle
• This is a series of events that prepares the cell for
division (Interphase) followed by actual process
of division (Mitosis)
• The cycle involves four stages;
– G1 Phase (rapid cell growth)
– S-Phase (DNA and Chromosomes replicates)
– G 2 Phase (mRNA’s & Proteins synthesis)
– M-Phase (Separation of replicated
chromosomes)
 Go Phase
• This is a sub phase of G1 in the cell cycle.
• This is also called resting phase, a period in which
cells exist in a quiescent state.
• As cells specialize they loose the ability to divide
i.e. they enter Go Phase, e.g. Nerve cells
• Cells in Go phase continue to perform their main
functions for the rest of an organisms life.
• However cells in Go can sometime re-enter the
cycle and proceed with cell division when need
arise. Eg. Myocytes when there is muscular injury
The circle represents the entire life cycle of the
cell, which can be divided into two major
phases:
1. interphase
2. division phase.
 Interphase
• Most of time spent by cell is in this phase
• Cells are not actively dividing
• It includes the G1, S and G2 phases
• Cells grow and undergo the various metabolic
processes needed for their functioning during
G1, S, and G2.
• Chromosomes are uncondensed throughout
interphase
 ‘

• G1 phase: cell undergo a period of rapid

growth, and the chromosomes are
unduplicated.

• S phase: cell begin to prepare for division

during interphase by duplicating its
chromosomes.

• G2 phase: G2, cell again grows and it

completes the preparations for division
(mitosis, or the M phase)
 ..

• In summary in interphase the cell replicates its

DNA and prepares for nuclear division.

• In humans, each of the 46 chromosomes

duplicates itself. The result is 46 duplicated
chromosomes, or 46 pairs of chromatids or
92 chromatids.
 The Stages of Mitosis and
Cytokinesis
• Mitosis has four stages named
1. Prophase
2. Metaphase
3. Anaphase
4. Telophase
• These stages are followed by the last cell
division named cytokinesis.
 Prophase
• Chromosomes in the nucleus become visible
under a microscope as they shorten and
thicken
• Centriole separates and its parts moves to the
opposite sides of the cell
• Centrioles with the spindle fibers forms the
spindle apparatus
 Metaphase
• Chromosomes composed of sister chromatids
move toward the center of the cell.

• This center area is called the equatorial plate,

as it is midway between the poles of the cell.

• Chromatids can become intertwined during

metaphase.
 Anaphase
• The centromeres divide and the sister
chromatids, now referred to as chromosomes,
move to opposite poles of the cell.
• If mitosis proceeds correctly, the same
number and type of chromosomes will be
found at each pole.
 Telophase
• The chromosomes reach the opposite poles of
the cell and begin to lengthen.

• The spindle fibres dissolve and a nuclear

membrane forms around each mass of
chromatin.

• Telophase is followed by cytokinesis, the

division of the cytoplasm.
 Cytokinesis
• Once the chromosomes have moved to
opposite poles, the cytoplasm begins to
divide.
• In an animal cell, a furrow develops, pinching
off the cell into two parts.
• This is the end of cell division
 Cell Cycle Checkpoints
• Cell cycle checkpoints are used by the cell to
monitor and regulate the progress of the cell
cycle
• There are three main checkpoints;
– G1/S checkpoint (Rate-limiting step/restriction point in the
cell cycle)
– G2/M checkpoint (Cell ensures that it has enough cytoplasm
and phospholipids for two daughter cells)
– Metaphase (mitotic) checkpoint (cell checks to ensure that
the spindle has formed and that all of the chromosomes are
aligned at the spindle equator before anaphase begins)
 .
• For single celled organisms, daughter cells are
“always” identical, unlike multicellular organism,
for instance stem cells which always undergoes
asymmetrical division.

• The process is highly regulated to prevents

imbalance and excessive growth, while ensuring
worn-out or damaged cells are replaced and
additional cells are formed in case of need eg
acclimatization in high altitude.
 Regulation of Cell cycle
• Cell replication is primarily controlled by
regulating the timing of nuclear DNA
replication and mitosis.

• The master controllers of these events are a

small number of heterodimeric protein kinases
that contain a regulatory subunit (cyclin) and
catalytic subunit (cyclin dependent kinase).
• The concentrations of the cyclins, the (regulatory
subunits) increase and decrease as cells progress
through the cell cycle.
• The catalytic subunits cyclin-dependent kinases
(CDKs), have no kinase activity unless they are
associated with a cyclin.
• Each CDK can associate with different cyclins, and
the associated cyclin determines which proteins
are phosphorylated by a particular cyclin-CDK
complex.
 Classes of Cyclins
• There are two main groups of cyclins:
• G1/S cyclins – essential for the control of the cell
cycle at the G1/S transition,
• Cyclin A – active in S phase.
• Cyclin D, and Cyclin E – regulates transition from G1 to S phase.
• G2/M cyclins – essential for the control of the cell
cycle at the G2/M transition.
• G2/M cyclins accumulate steadily during G2 and are
abruptly destroyed as cells exit from mitosis.
• Cyclin B  – regulates progression from G2 to M phase.
• Cyclin D and E are the mammalian G1 cyclins,
where Cyclin D is necessary for passage at
Restriction point.

• Cyclin A and B functions in the S phase, G2,

and early mitosis.
MEIOSIS
 MEIOSIS
• Is a specialized type of cell division that
generates haploid gametes from diploid
parental cell.

• Diploid (2n)  Haploid (n)

• It occurs only in specialized cells also known as

gonads (testes and ovaries)
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 Meiosis

• Meiosis is Two cell divisions

(called meiosis I and meiosis II)
• with only one duplication of chromosomes.
• Meiosis in males is called spermatogenesis
and produces sperm
• Meiosis in females is called oogenesis and
produces ova.
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 Steps in Meiosis
 Meiosis comprises two divisions: Meiosis I and II.
 Meiosis I is called reduction division, Meiosis II equitional division

Interphase I: G1, S and G2

Steps in Meiosis I:

Prophase I: Leptotene, Zygotene, Pachytene, Diplotene, Diakinesis

Metaphase I
Anaphase I
Telophase I
 Interphase I
• Similar to mitosis interphase.
• There is G1 phase, S phase and G2 phase
• In G1 phase, cell produces enzymes and structural
proteins needed for growth.
• In S phase Chromosomes replicate.
• G2 phase cell growth continues and proteins are
synthesized in preparation for mitosis
• Each duplicated chromosome consist of two identical
sister chromatids attached at their centromeres.
• Centriole pairs also replicate.

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46
 Prophase 1
Longest phase
Nucleus and nucleolus disappear

(a) Leptotene (b) Zygotene

• Individual • Chromosomes line
chromosomes coil (synapsis) up into
and condense into homologous
visible strands within chromosome pairs
the nucleus (bivalents or tetrads)
 (c) Pachytene
• Crossing over over a
small region of
homology between
nonsister chromatids
of homologous
chromosomes

• Chiasmata form

• Recombination of
information occurs
 Crossing Over
creates variation (diversity) in the offspring’s traits.

nonsister chromatids Tetrad

chiasmata: site of variation49

crossing over
 (d) Diplotene

• Homologous chromosomes separate

from each other a little
• In oogenesis, all oocytes develop to
this stage and are arrested
(dictyotene)
• Meiosis resumes to prepare oocyte
for ovulation at puberty or even later

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 (e) Diakinesis
• Chromosomes condense further and the four parts of the
tetrads are actually visible

• Chiasmata clearly visible

• Nucleoli disappear

• Nuclear membrane disintegrates into vesicles

• Meiotic spindle begins to form

• Two centrosomes containing a pair of centrioles migrate to the

two poles of the cell

• Microtubules attach to chromosomes at kinetochore

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Prophase I diagram
 Significance of crossing over
 Exchange of genetic materials between
maternal and paternal homologous
chromosomes
 Generates genetic variation
 Nondisjunction leading to trisomy or
monosomy
 Most monosomic or trisomic human embyos
are not viable
Metaphase I
• Homologous pairs align
along the metaphase
plate
• Tension is required to
balance bivalents along
the same equatorial line
• This necessitates at
least one cross over per
chromosome pair
• Hence independent
assortment occurs
Anaphase I
 Kinetochore MT
shorten
 Homologous
chomosomes pulled to
opposite poles
 Nonkitetochore MT
lengthen
 Centrosomes pushed
further apart
 Sister chromatids
remain attached
 Cell elongates
Telophase I
 Chromosomes arrive
at poles
 Each daughter cell is
haploid
 Each chromosome has
a pair of chromatids
 Spindle disappears
 Nuclear membrane
reforms
 Chromosomes uncoil
back into chromatin
 Cytokinesis
• Cell membrane
pinches

• Cell divides

• Formation of two
daughter cells
complete
 Meiosis II
• No interphase II
(or very short - no more DNA replication)

• Remember: Meiosis II is similar to mitosis

• No replication of DNA between meiosis I and

meiosis II

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 Prophase II
• same as prophase in mitosis
• Chromosomes condense and the spindle forms
• Nucleus and nucleolus disappear

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 Metaphase II
• Same as Metaphase in mitosis

• The chromosomes line up

individually on the equator of
the spindle
60
 Metaphase II

metaphase plate metaphase plate

61
 Anaphase II
• same as anaphase in mitosis

• sister chromatids separate

• At anaphase the centromeres divide, splitting the

two chromatids

• The one chromatid chromosomes are pulled to the

opposite poles
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 Anaphase II
• sister chromatids separate

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 Telophase II
• Same as telophase in mitosis.

• Nuclei form and nucleoli reform, spindle disappaers

• Cytokinesis occurs.

• Four haploid daughter cells produced, called

gametes ( sperms and eggs)

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Telophase II

65
 Meiosis II

Prophase II
Metaphase II
Anaphase II 4 Identical
Telophase II
haploid cells
Summary of Meiosis
RNA transcription
 Transcription
• Transcription is the first step of gene expression.
• During this process, the DNA sequence of a gene
is copied into RNA.
• Before transcription can take place, the DNA
double helix must unwind near the gene that is
getting transcribed.
• The region of opened-up DNA is called
a transcription bubble.
• Transcription uses one of the two exposed DNA
strands as a template; called the template strand.
• The RNA product is complementary to the
template strand and is almost identical to the
other DNA strand, called
the nontemplate (or coding) strand.
• There is one important difference: in the newly
made RNA, all of the T nucleotides are replaced
with U nucleotides.
• The site on the DNA from which the first RNA
nucleotide is transcribed is called the +1, 1 site,
or the initiation site.
• Nucleotides that come before the initiation site
are given negative numbers and said to
be upstream.
• Nucleotides that come after the initiation site
are marked with positive numbers and said to
be downstream.
 RNA Polymerase
• RNA polymerases are enzymes that transcribe
DNA into RNA.
• Using a DNA template, RNA polymerase builds
a new RNA molecule through base pairing.
• For instance, if there is a G in the DNA
template, RNA polymerase will add a C to the
new, growing RNA strand.
RNA polymerase always builds a new RNA strand in the 5’ to
3’ direction. That is, it can only add RNA nucleotides (A, U, C, or G) to
the 3' end of the strand.
• RNA polymerases are large enzymes with
multiple subunits, even in simple organisms
like bacteria.
• Humans and other eukaryotes have three
different kinds of RNA polymerases: I, II, and
III.
• Each one specializes in transcribing certain
classes of genes.
 Transcription Initiation
• To begin transcribing a gene, RNA polymerase
binds to the DNA of the gene at a region called
the promoter.
• The promoter tells the polymerase where to
"sit down" on the DNA and begin transcribing.
 Elongation
• Once RNA polymerase is in position at the promoter,
the next step of transcription—elongation—can begin.
• Elongation is the stage when the RNA strand
gets longer, due to the addition of new nucleotides.
• During elongation, RNA polymerase "walks" along one
strand of DNA, the template strand, in the 3' to 5'
direction.
• For each nucleotide in the template, RNA polymerase
adds a (complementary) sequence to the 3' end of the
RNA strand.
• The RNA transcript is nearly identical to
the non-template, or coding, strand of DNA.
• RNA strands have the base uracil (U) in place
of thymine (T), as well as a slightly different
sugar in the nucleotide.
• Each T of the coding strand is replaced with a
U in the RNA transcript.
 Termination
• RNA polymerase will keep transcribing until it
gets signals to stop.
• The process of ending transcription is
called termination.
• It happens once the polymerase transcribes a
sequence of DNA known as a terminator.
TRANSLATION
 Translation
• Translation is the RNA directed synthesis of
polypeptides
• The pathway of protein synthesis is called
translation because the ‘language’ of nucleotide
sequence on the mRNA is translated to the
‘language’ of amino acid sequence
• The mRNA is translated from its 5’ end to its 3’
end producing a protein synthesized from its
amino terminal end to its carboxyl terminal end
83
 Protein Synthesis
1. Activation of the amino acid
2. Initiation
3. Elongation
4. Termination and release
5. Folding and posttranslational processing

84
 Amino acid activation
Adding of Amino acid to tRNA
• Amino acid activation refers to the
attachment of an amino acid to its Transfer
RNA (tRNA).
• The  reactions are catalysed by a group of
enzymes called aminoacyl-tRNA synthetases
(named after the reaction product aminoacyl-
tRNA or aa-tRNA)
 Amino Acid Activation

tRNA
amino acid

adenylated amino acid

aminoacyl-
tRNA

AMP 86
 General Structure of
Aminoacyl-tRNA
• Aminoacyl group is
esterified to the 3’
position of the
terminal adenylate
residue
• Ester link
(highlighted):
activates amino acid
and links it to tRNA,
large free energy
release if hydrolysed

87
amino acid
(tryptophan) High
energy
tRNA bond
(tRNATrp)
High energy
bond

Linkage of amino acid

to tRNA
tRNA synthetase
(tryptophanyl tRNA
tRNA binds to its
synthetase)
codon in mRNA

base pairing

88
 Initiation
• When the mRNA interacts with the big
ribosome sub-unit, this triggers the approach
of transfer RNA (tRNA).
• The tRNA molecule possess a specific
sequence of 3-bases (anti-codon), which hast
to complement a corresponding sequence
(codon) within the mRNA sequence
 Initiation…
•  When it finds it, it attaches to the mRNA, as
the other end of the tRNA is “loaded” with an
amino acid.
• At this point arrives the other sub-unit of the
ribosome and a complete structure is formed.
• The first tRNA binds to a so called “start
codon”, which is one and the same for all
proteins
Translation Initiation complex
 Initiation of translation requires 4 specific steps;

1. A ribosome must dissociate into its' 40S and 60S

subunits.
2. A ternary complex termed the preinitiation
complex is formed consisting of the initiator, GTP,
eIF-2 and the 40S subunit
3. The mRNA is bound to the preinitiation complex
4. The 60S subunit associates with the preinitiation
complex to form the 80S initiation complex

92
 ELONGATION
• Elongation is a cyclic process on the
ribosome in which one amino acid at a
time is added to the nascent peptide chain

• The peptide sequence is determined by

the order of the codons in the mRNA
• Elongation involves several steps
catalyzed by proteins called elongation
factors (EFs)
• Each cycle uses four high-energy bonds (two from
the ATP used in amino acid activation to charge the
tRNA, and two from GTP)
08/10/2023 93
 Elongation involves 3 steps;

• Binding of aminoacyl-tRNA to the A site

• Peptide bond formation

• Translocation of the ribosome on the

mRNA

08/10/2023 94
 Binding of aminoacyl-tRNA to the A site
• In the initiation complex , both the A site (aminoacyl or acceptor
site) and E site (deacylated tRNA exit site) are free
• The binding of the appropriate aminoacyl-tRNA in the A site
requires proper codon recognition
• Elongation factor EF1A forms a ternary complex with GTP and
the entering aminoacyl -tRNA
• This complex then allows the correct aminoacyl-tRNA to enter the
A site with the release of EF1A GDP and phosphate.
• GTP hydrolysis is catalyzed by an active site on the ribosome;
hydrolysis induces a conformational change in the ribosome
concomitantly increasing affinity for the tRNA
• EF1A-GDP then recycles to EF1A-GTP with the aid of other
soluble protein factors and GTP
08/10/2023 95
 Peptide bond formation
• The -amino group of the new aminoacyl-tRNA in the A
site carries out a nucleophilic attack on the esterified
carboxyl group of the peptidyl-tRNA occupying the P site
(peptidyl site)
• This reaction is catalyzed by a peptidyltransferase, a
component of the 28S RNA of the 60S ribosomal subunit
• Because the amino acid on the aminoacyl-tRNA is
already "activated," no further energy source is required
for this reaction
• The reaction results in attachment of the growing peptide
chain to the tRNA in the A site

08/10/2023 96
 Translocation of the ribosome on mRNA

• The now deacylated tRNA is attached by its anticodon

to the P site at one end and by the open CCA tail to an
exit (E) site on the large ribosomal subunit
• At this point, elongation factor 2 (EF2 binds to and
displaces the peptidyl tRNA from the A site to the P site
• In turn, the deacylated tRNA is on the E site from
which it leaves the ribosome
• The EF2-GTP complex is hydrolyzed to EF2-GDP,
effectively moving the mRNA forward by one codon
and leaving the A site open for occupancy by another
ternary complex of amino acid tRNAEF1AGTP and
another
08/10/2023
cycle of elongation 97
 Ribosome Mechanism

Incoming aminoacyl-tRNA enters A-

site, base pairing with mRNA

High-energy bond of aminoacyl-

tRNA (3) broken and new bond
formed with (4), as the 2 tRNAs
shift to E- and P-sites
98
 mRNA moves 3
nucleotides through
ribosome, resetting it
and allowing the next
amino-acyl tRNA access
to A-site

And then it’s back to….

99
 TERMINATION
• Polypeptide chain termination occurs when a chain-
termination codon (stop codon) enters the A site of the
ribosome.
• The stop codons are UAA, UAG, and UGA.
• When a stop codon is encountered, a release factor
binds to the A site.
• A water molecule is added to the carboxyl terminus of
the nascent polypeptide, causing termination

08/10/2023 100
Protein chain released
into cytoplasm

mRNA
released and
ribosome
dissociates
into its 2
101
subunits
 Translation Summary
• tRNA is the adaptor between mRNA codon and amino acid
 It has complementary sequence to the codon (anticodon)
 The first base of the anti-codon may be a wobble base (does
not follow a watson crick base pairing)
 Aminoacyl tRNA synthetases activate amino acids by
attaching them to the appropriate tRNA

• The ribosome enables fast and accurate translation

 It has two subunits (large and small)
 It has three active sites (A, P, E)
 It has peptidyl transferase activity to promote peptide bond
formation (ribosome is a ribozyme)
 It coordinates a 3 step cycle of elongation
102
 Translation Summary
• Initiation factors enable initiation of translation
 Bacterial and eukaryotic initiation are very different in the
number of factors and how the start codon is defined/found
 Both form a preinititation complex to scan for the start codon
(AUG)
 Both position the small subunit on the mRNA 1st and use GTP
hydrolysis to load the large subunit

• Elongation factors make translation more efficient

 EF-Tu and EF-G shuttle in and out of the ribosome
 Both EF-Tu and EF-G use GTP hydrolysis to modulate the 3
step elongation cycle
 EF-Tu improves accuracy
 EG-G propels ribosome along mRNA
103
 Translation Summary
• Termination factors allow translation to stop
 There are two mechanisms at work in termination
 RF1/2 mimic tRNA and recognise the stop codons
 RF3 uses GTP hydrolysis

104