Protein synthesis and

regulation of genetic
information
 Protein synthesis
◼ The process of protein production.
◼ It actually consists of two processes — transcription and
translation.
◼ In eukaryotic cells, transcription takes place in the nucleus.
◼ During transcription, DNA is used as a template to make a
molecule of messenger RNA (mRNA).
◼ The molecule of mRNA then leaves the nucleus and goes
to ribosome in the cytoplasm, where translation occurs.
◼ During translation, the genetic code in mRNA is read and
used to make a protein.
These two processes are summed up by the central
dogma of molecular biology: DNA → RNA → Protein.
 Roles of RNA and DNA in
Protein synthesis
◼ DNA is the MASTER PLAN

◼ RNA is the BLUEPRINT of

the Master Plan

4
Just Like Construction…
◼ DNA is the master plan
◼ mRNA is the everyday blueprint
◼ rRNA is the builder
◼ tRNA is the gopher
◼ Amino Acids are the wood
◼ Proteins are the building
 Transcription
◼ Transcription is the first part of the central dogma
of molecular biology: DNA → RNA.
◼ Transcription is the synthesis of mRNA copied
from the DNA base sequences by RNA
polymerase.
◼ It is the transfer of genetic instructions in DNA to
mRNA.
◼ During transcription, a strand of mRNA is made
to complement a strand of DNA.
 Three Steps of Transcription
◼ 1. Initiation is the beginning of transcription.
 It occurs when the enzyme RNA polymerase binds to a region of
a gene called the promoter.
 This signals the DNA to unwind so the enzyme can “read” the
bases in one of the DNA strands.
 The enzyme is ready to make a strand of mRNA with a
complementary sequence of bases.

◼ 2. Elongation is the addition of nucleotides to the mRNA

strand.

◼ 3. Termination is the ending of transcription. The mRNA

strand is complete, and it detaches from DNA.
 Processing mRNA
◼ In eukaryotes, the new mRNA is not yet ready
for translation.
◼ At this stage, it is called pre-mRNA, and it must
go through more processing before it leaves the
nucleus as mature mRNA.
◼ The processing may include splicing, editing,
and polyadenylation.
◼ These processes modify the mRNA in various
ways.
◼ Such modifications allow a single gene to be
used to make more than one protein.
Splicing
◼ Splicing removes introns from mRNA.
 Introns are regions that do not code for the protein.

◼ The remaining mRNA consists only of regions

called exons that do code for the protein.

◼ The ribonucleoproteins are small proteins in the

nucleus that contain RNA and are needed for
the splicing process.
Editing
◼ Editing changes some of the nucleotides in
mRNA.

◼ For example, a human protein called APOB,

which helps transport lipids in the blood, has two
different forms because of editing.

◼ One form is smaller than the other because

editing adds an earlier stop signal in mRNA.
Polyadenylation
◼ Polyadenylation adds a “tail” to the mRNA.
◼ The tail consists of a string of A’s (adenine bases).
◼ It signals the end of mRNA.
◼ It is also involved in exporting mRNA from the
nucleus, and it protects mRNA from enzymes that
might break it down.
◼ May help in initiation of translation.
Differences between prokaryotes and
eukaryotes in transcription
Prokaryotic Transcription Eukaryotic Transcription
Transcription and translation occur Transcription and translation don’t
simultaneously occur simultaneously.

Prokaryotic transcription occurs in the Eukaryotic transcription occurs in the

cytoplasm nucleus and translation occurs in the
cytoplasm.

RNAs are released and processed in RNAs are released and processed in
the cytoplasm the nucleus

RNA polymerases are a complex of RNA polymerases are a complex of

five polypeptides. 10 -15 polypeptides.

Doesn’t require any proteins or other Requires proteins known as

factors for the initiation of transcription factors for the initiation.
transcription
 Translation
◼ Translation is the second part of the central
dogma of molecular biology: RNA → Protein.
◼ It is the process in which the genetic code in
mRNA is read to make a protein.
◼ After mRNA leaves the nucleus, it moves to a
ribosome, which consists of rRNA and proteins.
◼ The ribosome reads the sequence of codons in
mRNA, and molecules of tRNA bring amino
acids to the ribosome in the correct sequence.
Cont.
◼ To understand the role of tRNA, you need to
know more about its structure.
◼ Each tRNA molecule has an anticodon for the
amino acid it carries.
◼ An anticodon is complementary to the codon for
an amino acid.
◼ Their function is to base pair with the codon on a
strand of mRNA during translation.
◼ For example, the amino acid lysine has the
codon AAG, so the anticodon is UUC.
◼ Therefore, lysine would be carried by a tRNA
molecule with the anticodon UUC.
The anticodon sequence determines the
amino acid that the tRNA carries.
Cont.
◼ Wherever the codon AAG appears in mRNA, a
UUC anticodon of tRNA temporarily binds.
◼ While bound to mRNA, tRNA gives up its amino
acid.
◼ With the help of rRNA, bonds form between the
amino acids as they are brought one by one to
the ribosome, creating a polypeptide chain.
◼ The chain of amino acids keeps growing until a
stop codon is reached.
Protein Synthesis: Translation
 This is a molecule of messenger RNA.
mRNA is transcribed in the nucleus.

codon
A U G G G C U U AAA G C A G U G C A C G U U

mRNA molecule
 A ribosome attaches to the mRNA
molecule.

ribosome

A U G G G C U U AAA G C A G U G C A C G U U
 Amino acid

tRNA molecule

A transfer RNA molecule arrives.

It brings a specific amino acid to the first
three bases (codon) on the mRNA.
anticodon The three unpaired bases (anticodon)
on the tRNA link up with the codon.
UAC
A U G G G C U U AAA G C A G U G C A C G U U
 Another tRNA molecule comes into
place, bringing a second amino acid.

Its anticodon links up with the second

codon on the mRNA.

UAC
A U G G G C U U AAA G C A G U G C A C G U U
 Peptide bond

A peptide bond forms between the

two amino acids.

A U G G G C U U AAA G C A G U G C A C G U U
 The first tRNA molecule releases its amino
acid and moves off into the cytoplasm.

A U G G G C U U AAA G C A G U G C A C G U U
 The ribosome moves along the mRNA to
the next codon.

A U G G G C U U AAA G C A G U G C A C G U U
 Another tRNA molecule brings
the next amino acid into place.

A U G G G C U U AAA G C A G U G C A C G U U
 A peptide bond joins the second
and third amino acids to form a
polypeptide chain.

A U G G G C U U AAA G C A G U G C A C G U U
The process continues.
The polypeptide chain gets longer.

This continues until a termination

(stop) codon is reached.

The polypeptide is then complete.

A U G G G C U U AAA G C A G U G C A C G U U
 mRNA coding region
◼ Each mRNA strand has a coding region within it that codes
for protein synthesis.
◼ The coding region starts with the AUG start, and continues
with the established reading frame.
◼ The coding region ends when a stop codon is reached.
◼ The mRNA strand prior to the start codon is called the 5’
untranslated region or leader sequence.
◼ The mRNA strand after the stop codon is called the 3’
untranslated region or trailing sequence.
◼ Collectively, the leader sequence and trailing sequence are
referred to as noncoding regions of the mRNA.
.
 Gene regulation
◼ Refers to the mechanisms that act to induce or
repress the expression of a gene.

◼ These include structural and chemical changes

to the genetic material, binding of proteins to
specific DNA elements to regulate transcription,
or mechanisms that modulate translation of
mRNA.
Gene regulation
 Positive Regulation
• Positive regulation involves activators
• Enhance activity of RNA polymerase

• Activator-binding
sites are near
promoters that
weakly bind RNA Pol
or do not bind at all.
• It may remain bound
until a molecule
signals dissociation.
• Alternatively, the
activator may only
bind when signaled.
 Negative Regulation
◼ Negative regulation involves repressors
 Example: Repressor binds to DNA and shuts down
transcription
 Alternative: Signal causes repressor to dissociate
from DNA; transcription induced

Despite opposite effects

on transcription, both
are negative regulation
 Regulation of gene expression in
eukaryotes
◼ Levels of control of gene expression can be:
 Short term control – to meet the daily needs of the organism.
 Long term control – gene regulation in
development/differentiation.
◼ Prokaryote gene expression typically is regulated by an
operon, the collection of controlling sites adjacent to
polycistronic protein-coding sequences.
◼ Eukaryotic genes are regulated in units of protein-coding
sequences and adjacent controlling sites, but operons are
not known to occur.
◼ Eukaryotic gene regulation is more complex because
eukaryotes possess a nucleus.
Operon
◼ An operon typically includes:
 Regulator gene – this codes for a DNA-binding
protein that acts as a repressor.
 Promoter – DNA sequence that binds RNA
polymerase.
 Operator – portion of DNA where an active
repressor binds.
 Structural genes – codes for enzymes and proteins
needed for the operons metabolic pathway.
Prokaryotic cells turn genes on and off by controlling transcription.
▪ Gene expression is the
activation of a gene that
results in transcription and
the production of mRNA.
▪ A promoter is the segment of
DNA that is recognized by the
enzyme RNA polymerase,
which then initiate
transcription.
▪ An operator is the segment of
DNA that acts as a switch by
controlling the access of RNA
polymerase to structural gene.
▪ In prokaryotes, the structural
genes, the promoter and
operators collectively form an
operon.
 Two operons of E. Coli:
◼ trp operon
 Regulates the expression of tryptophan.
 Normally “on” so it expresses the structural genes
needed.
 If tryptophan is already present, it binds to the repressor
causing it to change shapes so it can bind to the
promoter.
◼ lac operon
 Regulation for lactose metabolism.
 The repressor is normally bound to the operator to turn
“off” gene expression.
 In the presence of lactose, lactose binds to the repressor
causing it to change shape so DNA polymerase can
easily bind to the promotor.
Eukaryote gene expression is regulated at six levels:

1. Transcription control

2. RNA processing

3. mRNA transport

4. mRNA translation

5. mRNA degradation

6. Protein degradation
1. Transcription control controlled by:

a. Promoters
▪ Occur upstream of the transcription start site.
▪ Some determine where transcription begins (e.g.,
TATA), whereas others determine if transcription
begins.
▪ Promoters are activated by specialized transcription
factor (TF) proteins (specific TFs bind specific
promoters).
▪ One or many promoters (each with specific TF
proteins) may occur for any given gene.
▪ Promoters may be positively or negatively regulated.
1. Transcription control controlled by:
b. Enhancers

▪ Occur upstream or downstream of the transcription

start site.

▪ Regulatory proteins bind specific enhancer

sequences; binding is determined by the DNA
sequence.

▪ Loops may form in DNA bound to TFs and make

contact with upstream enhancer elements.

▪ Interactions of regulatory proteins determine if

transcription is activated or repressed (positively or
negatively regulated).
2. RNA processing control

• RNA processing regulates mRNA production from

precursor RNAs.
• Two independent regulatory mechanisms occur:

• Alternative polyadenylation = where the polyA tail is

added
• Alternative splicing = which exons are spliced

• Alternative polyadenylation and splicing can occur

together.
• Example: Sex determination in Drosophila
3. mRNA transport control

▪ Eukaryote mRNA transport is regulated.

▪ Some experiments show ~1/2 of primary transcripts never

leave the nucleus and are degraded.

▪ Mature mRNAs exit through the nuclear pores.

4. mRNA translation control
▪ Unfertilized eggs are an example, in which mRNAs
(stored in the egg/no new mRNA synthesis) show
increased translation after fertilization).

▪ Presence or absence of the 5’ cap and the length of the

poly-A tail at the 3’ end can determine whether translation
takes place and how long the mRNA is active.

▪ Conditions that affect the length of the poly-A tail or leads

to the removal of the cap may trigger the destruction of an
mRNA.
5. mRNA degradation control
▪ All RNAs in the cytoplasm are subject to degradation.

▪ tRNAs and rRNAs usually are very stable; mRNAs vary

considerably (minutes to months).

▪ Stability may change in response to regulatory signals and

is thought to be a major regulatory control point.
6. Post-translational control-protein degradation
▪ Proteins can be short-lived (e.g., steroid receptors) or
long-lived (e.g., lens proteins in your eyes).
▪ This is a cells last change to affect gene expression.
▪ Protease, enzymes that break down proteins, are confined
to lysosomes and proteasomes.
▪ When a protein is tagged by a signaling protein, it enters a
proteasome to be degraded.
 MUTATIONS
❖ Alterations in existing genes.
❖ Forms of mutation:
1. Gene mutations – (Point mutation &
frameshift) - single DNA codon
Structural changes such as deletion, inversion,
insertion of LINES or SINES, transposition, etc
of a larger portion of DNA.

2. Chromosomal mutations - structural

modifications in chromosomes and even missing or
extra chromosomes.
 A. Point mutation (substitution)
◼ It is a mutation that exchange one base for
another. Forms/Types of substitution include:
 Missense mutation:
◼ For example, CTC in the DNA sense strand [GAG in mRNA]
will specify a glutamate residue in the protein; this is altered to
CAC in the DNA or GUG in the mRNA, resulting in a valine
residue in the β-globin protein chain causing sickle-cell
anaemia
 Silent mutation:
◼ wobble" base pairing (G-U), (I-U), (I-A), (I-C)
 Nonsense mutation:
◼ a stop codon and cause a truncated protein.
 B. Frameshift mutation

◼ Insertion
 extra base pairs are inserted into a new place in the
DNA

◼ Deletions
 2. Chromosomal mutation
1. Deletion
2. Duplication
3. Inversion
4. Translocation
 Agents that Damage DNA
1. Radiations
 Highly reactive oxygen radicals produced during
normal cellular respiration as well as by other
biochemical pathways.

 Ionizing radiation such as gamma rays and x-rays.

 Ultraviolet rays, especially the UV-C rays (~260 nm)

that are absorbed strongly by DNA but also the
longer-wavelength UV-B that penetrates the ozone
shield.
 Agents that Damage DNA
2. Chemicals in the environment
 Aromatic hydrocarbons, including some found in
cigarette smoke.

 Plantand microbial products, e.g. the Aflatoxin

produced in moldy peanuts.

 Chemicalsused in chemotherapy, especially

chemotherapy of cancers.
 Agents that Damage DNA
3. Free radicals
◼ Oxidative damage mainly occurs by formation of
free radical or radiomimetic compounds.
 70% damage by OH-
 Radiolysis of H2O produce peroxides

 Formation of OH, requires metal near to the

DNA
 Fe2+ mainly forms OH, formation by fenton
reaction Fe² + H2O2 → Fe³3+ + OH· + OH−
 GROUP ASSIGNMENT
1. Describe the genetic code and explain why it is
considered almost universal.
2. Discuss sex determination in fish.
3. Describe mechanisms of epigenetic regulation in
eukaryotes.
4. Explain environmental factors as the influence of
phenotypic variation.
5. How a genetic bottleneck reduces genetic
diversity?
 GROUP ASSIGNMENT
6. Why errors in DNA replication can be far more
damaging than errors in transcription?
7. Write notes on Genetically Improved Farmed Tilapia
(GIFT).
8. The pros and cons of crossbreeding and hybridization
in aquaculture.
9. What are the advantages of cross-breeding Nile tilapia
and Oreochromis spilurus?
 GROUP ASSIGNMENT
Hints:
◼ 1. Each group should attempt one question.

◼ 2. PPT should not be more than 10 slides.

◼ 3. Submission of word file and presentation

will be on 27th January 2025.

◼ 4. Word file and PPT should be different.

 END

Thanks for your attention