Translation

Tigist T.Bekele 1
Changing nucleic acids into amino acids
The three-base code in DNA or mRNA is called a
codon.
And the codon from mRNA become decoded by
the tRNA in the ribosome. The anti-codons in
tRNA corresponds with one amino acid
• This is where TRANSLATION begins

Tigist T.Bekele 2
 TRANSLATION

 Translation begins when mRNA binds to the

RIBOSOME in the cell.
 In translation, tRNA molecules act as the
interpreters of the mRNA codon sequence.
 At the middle of the folded strand, there is a
three-base coding sequence in the tRNA called
the anticodon.
 Each anti-codon is complementary to a codon
on the mRNA.

Tigist T.Bekele 3
 The role of Ribosomes
The third type of RNA is
ribosomal RNA (rRNA).
Ribosomes are made of
rRNA and PROTEIN.
Ribosomes are the
‘decoding’ units of the cell.
(Sites of protein synthesis) P A
Ribosomes consist of two
major components — the
small ribosomal subunit
which reads the RNA, and
the large subunit which
joins amino acids to form a
polypeptide chain. Tigist T.Bekele 5
 Messenger RNA
(mRNA)

mRNA: Composed of Codons

Codons are 3-base sequences of mRNA
Messenger RNA (mRNA)
Ribosomes
Transfer RNA (tRNA)
Amino Acids
Tigist T.Bekele 6
 Ribosomes

Made of rRNA and protein

2 subunits (large and small) form a 3D groove
2 major sites: P site---holds the growing polypeptide
A site---new amino acids enter here

Tigist T.Bekele 7
 Step 1: Initiation
5-cap of mRNA binds to ribosome
Start codon AUG and anticodon with
Methionine bind a P site-starts translation
A site is open and ready to receive new tRNAs

Tigist T.Bekele 8
Step 2: Elongation
Codon recognition
Translocation: ribosome
moves along mRNA,
aminoacyl tRNA shifts
from A site to P site
Peptide bond formation

Tigist T.Bekele 9
Tigist T.Bekele 10
The Genetic Code
• There are 64
(4X4X4) possible
triplet codes, but
only 20 amino
acids.
• As seen in the
table, more than 1
triplet may code for
the same amino
acid.
• Note that several
codons can also act
as start codons Tigist T.Bekele 11
Tigist T.Bekele 12
 Step 3:
Termination
A stop codon
is reached-
translation
stops
UAA UAG
UGA
All parts
release

Tigist T.Bekele 13
 The three codons UAA, UAG and UGA do not code
for amino acids. They act as stop signals in protein
synthesis. These three codons are collectively
known as termination codons or non-sense codons.
The codons UAG, UAA and UGA are often referred
to, respectively, as amber and opal codons.
Other characteristics of genetic code
The genetic code is
• Universal
• Specific
• Nonoverlapping
• Degenerate/redundant
The codons that designate the same amino acid are called
synonyms. Most of the synonyms
Tigist T.Bekele differ only in the third
14 (3'
 Translation Polypeptides and Mutation

Normally, the genetic code is

translated and the correct protein is
formed from a long chain of amino
acids.
Mutation
Any change in the nucleotide
sequence of DNA
Mutations can involve large sections
of chromosomes or single base pairs
Tigist T.Bekele 15
Deletion or insertion mutations are
most disruptive because they change
the reading frame, causing a frame
shift.
Substitution mutations have varied
impact on amino acid sequences.
Substitutions of 1st or 2nd base in
codon almost always changes the
amino acid
Substitution of 3rd base in codon does
not always change the amino acid
Tigist T.Bekele 16
Tigist T.Bekele 17
 Sickle Cell Anemia
Normal Hemoglobin Sickle Cell Hemoglobin
DNA GGA CTT GCA GGA CAT GCA
mRNA CCU GAA CGU CCU GUA CGU
A.A. PRO GLU ARG PRO VAL ARG

Changes in one or a few bases is called a Point

Mutation
2 Types: Substitution
Insertion/Deletions

Tigist T.Bekele 18
Translation Polypeptides and Mutation
Sickle cell “Normal” red blood
cell

Tigist T.Bekele 19
Gene Expression & Regulation of
gene Expression

1
 Gene
Expression
• It is the process by which information
from a gene is used in the synthesis of a
functional gene product.

• These products are often proteins, but

in non-protein coding genes such as
rRNA genes
or tRNA genes, the product is a
functional RNA
Gene expression regulation:
Both of these cells contain the same
genome, but they express different
RNAs and proteins.
Tigist T.Bekele 23
Classification of gene with respect to their
Expression:
Constitutive ( house keeping) genes:
1. Are expressed at a fixed rate, irrespective to
the cell condition.
2 Their structure is simpler

Controllable genes:
3. Are expressed only as needed. Their amount
may increase or decrease with respect to different
condition.
4.Their structure is relatively complicated with
some response elements
 1. positive
regulation:

• When the expression of gene is

quantitatively increased by the
presence of specific regulatory
element is known as positive
regulation.
• An element modulating a positive
regulation is known as
activator or positive
regulator.
Tigist T.Bekele 25
 2. Negative
regulation.
• when the expression of genetic
information is diminished by the
presence of specific regulatory element.

• The element or molecule mediating the

negative regulation is said to be repressor.

Tigist T.Bekele 26
Levels of regulation of gene
expression

.
Purpose of regulation of gene
expression
Regulated expression of genes is required for
1) Adaptation- Cells of multicellular
organisms respond to varying conditions.
Such cells, exposed to hormones and growth
factors, change substantially in –
o shape,

o growth rate, and

o other characteristics
.
 Purpose of regulation of gene
expression
2) Tissue specific differentiation and development
• The genetic information present in each somatic cell
of a organism is practically identical.
• Cells from muscle and nerve tissue show strikingly
different morphologies and other properties, yet
they contain exactly the same DNA.
• These diverse properties are the result of
differences in gene expression.

.
 Differences between gene
expression in prokaryotes
and eukaryotes
Gene regulation is significantly more
complex in eukaryotes than in prokaryotes
for a number of reasons:
1) First, the genome being regulated is
significantly larger
• Human cell have larger genome

.
 2) Different cell types

• Different cell types are present in most

eukaryotes.
• Liver and pancreatic cells, for example, differ
completely in the genes that are expressed.
• Different mechanisms are involved in the
regulation of such genes.

.
 3) Absence of operons

• The eukaryotic genes are not generally

organized into operons as are there in
prokaryotes

• Instead, genes that encode proteins for steps

within a given pathway are often spread
widely across the genome.
.
Tigist T.Bekele 33
4) Chromatin structure
• The DNA in eukaryotic cells is
extensively folded and packed into the
protein-DNA complex called
chromatin.
• Histones are an important part of this
complex since they both form the
structures known as nucleosomes and also
contribute significantly into gene
regulatory mechanisms.
.
Gene expression in
Eukaryotes
The genetic constitutions of nearly all somatic cells
are identical.
Tissue or cell specificity is dictated by differences in
gene expression of this complement of genes.
Alterations in gene expression allow a cell to adapt to
environmental changes.
Gene expression can be controlled at multiple
levels by chromatin modifications, changes in
transcription, RNA processing, localization, and
stability or utilization.
Gene amplification and rearrangements also
influence gene expression. .
Eukaryotic gene regulation occurs
at several levels:
G.E control at DNA
level
Is done by
• Gene amplification
• Gene rearrangement
 Gene Amplification

• The gene product can be increased by

increasing the number of genes available for
transcription of specific molecules
• Among the repetitive DNA sequences are
hundreds of copies of ribosomal RNA genes
and tRNA genes.

,
 Gene Amplification
(contd.)
• Such requirements are fulfilled by
amplification of these specific genes.
• Subsequently, these amplified genes,
presumably generated by a process of
repeated initiations during DNA
synthesis, provide multiple sites for
gene transcription.

.s
Gene rearrangement is observed during
immunoglobulins synthesis.

here segment of DNA move from one location to

another in the genom.These DNA coding changes
are needed for generating the required recognition
diversity central to appropriate immune function.
.
 G.E control-Transcriptional level

1) Chromatin Remodeling
• Large regions of chromatin are
transcriptionally inactive /heterochromatin in
some cells while they are either active or
potentially active /euchromatin in other
specialized cells

For example, the DNA containing the -globin

gene cluster is "active" chromatin in the
.
2) Use of alternative promoter
Some genes has series of promoter's showing
tissue specific expression. So in different
tissue, different transcript and different
protein produced
3) Histone acetylation and
deacetylation
• Acetylation is known to occur on lysine
residues in the amino terminal tails of
histone molecules.
• This modification reduces the positive
charge of these tails and decreases
the binding affinity of histone for the
negatively charged DNA.
• Accordingly, the acetylation of histones
could result in disruption of nucleosomal
structure and allow readier access of
4) DNA
methylation
is :
the addition or removal of a methyl group
predominantely where cytosine bases occur
consecutively.
Post transcriptional regulation of
gene expression
Done by
• Alternative splicing
• Class switching
• Regulation of RNA
stability
• RNA editing
 Alternative RNA
Processing
• This can result when alternative
promoters, intron-exon splice sites,
or polyadenylation sites are used.

• Occasionally, heterogeneity within a cell

results, but more commonly the same
primary transcript is processed
differently in different tissues.
.
 Alternative RNA Processing
(contd.)
Alternative splicing and processing,
results in the formation of seven unique -
tropomyosin mRNAs in seven different
tissues.
 Class
switching

• In this process one

gene is switched
off and a closely
related gene
takes up the
function.
• During intrauterine
life embryonic Hb is
the first Hb to be
formed.
 mRNA stability
• Changes in the stability of a specific mRNA
can therefore have major effects on biologic
processes.
• The stability of the mRNA can be
influenced by hormones and certain other
effectors.
• The ends of mRNA molecules are
involved in mRNA stability.
• The 5' cap structure in eukaryotic mRNA
prevents attack by 5' exonucleases, and the
.
RNA Editing
Tigist T.Bekele 53
Post translational
regulation
• Protein activation
• proteins are not active when first formed. so
to be active they undergo modifications
Folding,
Proteolysis
Enzymatic
cleavage
Regulation of
protein