PROTEIN SYNTHESIS:

TRANSLATION OF THE
GENETIC MESSAGE

SUBMITTED BY: GROUP 10

DAGAMI, RICHEL
GALAN, RONALIZA
LERIOS, MARIANNE ANGELI
MARTIJA, CHERRY KAY
SEROJIHOS, JOY
TRANSLATING THE
GENETIC MESSAGE
 TRANSLATING THE GENETIC
MESSAGE

 Protein biosynthesis is a complex process requiring

ribosomes, messenger RNA (mRNA), transfer RNA
(tRNA), and a number of protein factors.
 The ribosome is the site of protein synthesis.
 The mRNA and tRNA, which are bound to the
ribosome in the course of protein synthesis, are
responsible for the correct order of amino acids in the
growing protein chain.
 TRANSLATING THE GENETIC
MESSAGE
The formation of a protein takes place in four steps:
 
1. Activation

2. Initiation

3. Elongation

4. Termination
TRANSLATING THE GENETIC
MESSAGE
STEPS IN PROTEIN BIOSYNTHESIS
THE GENETIC CODE
 THE GENETIC CODE

The most important features of the code can be specified by

saying that the genetic message is contained in a:

1. Triplet

2. Nonoverlapping

3. Commaless

4. Degenerate

5. Universal code
 THE GENETIC CODE

1. A triplet code means that a sequence of three bases

(called a codon) is needed to specify one amino acid.

2. The term nonoverlapping indicates that no bases are

share between consecutive codons; the ribosome
moves along the mRNA three bases at a time rather
than one or two at a time. If the ribosome moved along
the mRNA more than three bases at a time, this
situation would be referred to as “a punctuated code.”
 THE GENETIC CODE

THEORETICALLY
POSSIBLE
GENETIC CODE
 THE GENETIC CODE

3. No intervening bases exist between codons, the code is

commaless.

4. In a degenerate code, more than one triplet can encode

the same amino acid. Sixty-four (4 4 4) possible triplets
of the four bases occur in RNA, and all are used to
encode the 20 amino acids or one of the three stop
signals.
 THE GENETIC CODE

HOW DID SCIENTIST

DETERMINE THE GENETIC
CODE?
 THE GENETIC CODE

A variety of techniques were used to determine the

genetic code. The most comprehensive experiment was
the filter-binding experiment of Nirenberg, which
used specific trinucleotides bound to a filter to see
which tRNA molecules would bind.
 THE GENETIC CODE

The filter-binding assay

for elucidation of the
genetic code.
 THE GENETIC CODE

 CODON-ANTICODON PAIRING AND WOBBLE

- A codon forms base pairs with a complementary

anticodon of a tRNA when an amino acid is
incorporated during protein synthesis. Because there are
64 possible codons, one might expect to fi nd 64 types
of tRNA but, in fact, the number is less than 64 in all
cells.
 THE GENETIC CODE

IF THERE ARE 64 CODONS,

HOW CAN THERE BE LESS
THAN 64 tRNA MOLECULES?
 THE GENETIC CODE

There are only about half as many tRNA molecules as

there are codons for amino acids. This occurs because of
wobble in the third base (5' end) of the anticodon on the
tRNA. The wobble base can break Watson– Crick base-
pairing rules under certain circumstances. For example,
if the wobble base is a U, it can bind to an A or a G of
the mRNA codon, and thus fewer tRNA molecules are
needed.
 THE GENETIC CODE

“Wobble” base pairing.

THE GENETIC CODE

Various base-pairing alternatives

AMINO ACID ACTIVATION
AMINO ACID ACTIVATION

▪ The activation of the amino acid and the formation

of the aminoacyl-tRNA take place in two separate
steps, both of which are catalyzed by the
aminoacyltRNA synthetase. First, the amino acid
forms a covalent bond to an adenine nucleotide,
producing an aminoacyl-AMP. The free energy of
hydrolysis of ATP provides energy for bond
formation. The aminoacyl moiety is then
transferred to tRNA, forming an aminoacyl-tRNA.
 AMINO ACID ACTIVATION

The aminoacyl-
tRNA synthetase
reaction
AMINO ACID ACTIVATION

WHAT IS THE “SECOND

GENETIC CODE”?
 AMINO ACID ACTIVATION

The second genetic code refers to the specificity with

which the aminoacyltRNA synthetases put the correct
amino acid onto the correct tRNA. There is only
minimal proofreading past that point, so the correct
loading of amino acids is critical. If the wrong amino
acid is loaded on the tRNA, it is quickly hydrolyzed by
the synthetase to avoid such errors.
 AMINO ACID ACTIVATION

Ribbon diagram of the

tRNA tertiary
structure
PROKARYOTIC
TRANSLATION
PROKARYOTIC TRANSLATION

 RIBOSOMAL ARCHITECTURE

- Protein synthesis requires the specifi c binding of

mRNA and aminoacyl-tRNAs to the ribosome.
Ribosomes have a specifi c architecture that
facilitates the binding.
PROKARYOTIC TRANSLATION

 CHAIN INITIATION

- In all organisms, the synthesis of polypeptide chains

starts at the N-terminal end; the chain grows from
the N-terminal end to the C-terminal end. The
coding strand sequences are read from 5' to 3', the
mRNA is read from 5' to 3', and the proteins are
built from the N-terminus to the C-terminus. In
prokaryotes, the initial N-terminal amino acid of all
proteins is N-formylmethionine (fmet) .
PROKARYOTIC TRANSLATION

Formation of the N-formylmethionine-

tRNAfmet (first reaction)
PROKARYOTIC TRANSLATION

HOW DOES THE RIBOSOMES

KNOW WHERE TO START
TRANSLATING?
PROKARYOTIC TRANSLATION

• For the mRNA to be translated correctly, the ribosome

must be placed at the correct start location.

• The start signal is preceded by a purine-rich leader

segment of mRNA, called the Shine–Dalgarno
sequence (5'-GGAGGU-3') 90410_12_c12_p331-
362.indd 342 10/18/07 5:51:03 PM 10/18/07 5:51:03
PM which usually lies about 10 nucleotides upstream
of the AUG start signal (also known as the initiation
codon) and acts as a ribosomal binding site.
PROKARYOTIC TRANSLATION

The formation of an
initiation complex
PROKARYOTIC TRANSLATION

 CHAIN ELONGATION

- The elongation phase of prokaryotic protein

synthesis uses the fact that three binding sites for
tRNA are present on the 50S subunit of the 70S
ribosome. The three tRNA binding sites are called
the P (peptidyl) site, the A (aminoacyl) site, and the
E (exit) site.
PROKARYOTIC TRANSLATION

A summary of the steps

in chain elongation
PROKARYOTIC TRANSLATION

Why is EF-Tu so important in E.

coli?
PROKARYOTIC TRANSLATION

The elongation factors EF-Tu and EF-Ts add a level of

complexity to the elongation of proteins. This
complexity seems at first to be burdensome, but is
actually very important as it leads to another level of
proofreading. EF-Tu delivers the aminoacyl-tRNA to
the A site of the ribosome only when the codon and
anticodon match.
PROKARYOTIC TRANSLATION

 CHAIN TERMINATION

- A stop signal is required for the termination of

protein synthesis. The codons UAA, UAG, and
UGA are the stop signals. These codons are not
recognized by any tRNAs, but they are recognized
by proteins called release factors.
PROKARYOTIC TRANSLATION

The events in peptide

chain termination
PROKARYOTIC TRANSLATION

 THE RIBOSOMES IS A RYBOZYME

Until recently, proteins were thought to be the only

molecules with catalytic ability. Then the self-splicing
ability of the Tetrahymena snRNP showed that RNA
can also catalyze reactions. In 2000, the complete
structure of the large ribosomal subunit was determined
by X-ray crystallography to 2.4-Å (0.24-nm) resolution
PROKARYOTIC TRANSLATION

Ribosome structure
PROKARYOTIC TRANSLATION

 POLYSOMES

- complex of mRNA with several ribosomes is

called a polysome; an alternative name is
polyribosome.
PROKARYOTIC TRANSLATION

Simultaneous protein synthesis on polysomes

PROKARYOTIC TRANSLATION

 COUPLED TRANSLATION

- prokaryotic gene is being simultaneously

transcribed and translated.
PROKARYOTIC TRANSLATION

Electron micrograph
showing coupled
translation
EUKARYOTIC
TRANSLATION
 EUKARYOTIC TRANSLATION

 The main features of translation are the same in

prokaryotes and eukaryotes, but the details differ. The
messenger RNAs of eukaryotes are characterized by
two major posttranscriptional modifi cations. The fi rst
is the 5' cap, and the second is the 3' poly A tail Both
modifi cations are essential to eukaryotic translation.
EUKARYOTIC TRANSLATION

How is translation different in

eukaryotes?
 EUKARYOTIC TRANSLATION

• Eukaryotic mRNA undergoes a lot of processing that is

not observed in prokaryotes.

• Eukaryotic mRNA has a 5' cap and a 3' poly A tail,

both of which are involved in formation of the
initiation complex.

• There is no Shine–Dalgarno sequence, but there is a

Kozak sequence surrounding the correct AUG start
codon.
 EUKARYOTIC TRANSLATION

• The number of initiation factors and elongation factors

is much higher in eukaryotes than in prokaryotes.

• Stop codons in eukaryotes are sometimes recognized

by suppressor tRNAs, which allow insertion of
nonstandard amino acids, such as selenocysteine.
 EUKARYOTIC TRANSLATION

 Chain Initiation

- Thirteen more initiation factors are given the

designation eIF, for eukaryotic initiation factor.
Many of them are multisubunit proteins.
 EUKARYOTIC TRANSLATION

The three stages in the

initiation of
translation in
eukaryotic cells
 EUKARYOTIC TRANSLATION

 Chain Elongation

• Peptide chain elongation in eukaryotes is very similar

to that of prokaryotes.

• same mechanism of peptidyl transferase and ribosome

translocation is seen.

• structure of the eukaryotic ribosome is different in that

there is no E site, only the A and P sites.
EUKARYOTIC TRANSLATION

• There are two eukaryotic elongation factors, eEF1

and eEF2.

• . The eEF1 consists of two subunits, eEF1A and

eEF1B. The 1A subunit is the counterpart of EF-Tu in
prokaryotes. The 1B subunit is the equivalent of the
EF-Ts in prokaryotes.

• The eEF2 protein is the counterpart of the

prokaryotic EF-G, which causes translocation.
 EUKARYOTIC TRANSLATION

 Chain Termination

• In eukaryotes, only one release factor binds to all

three stop codons and catalyzes the hydrolysis of the
bond between the C-terminal amino acid and the Trna

• There is a special tRNA called a suppressor tRNA,

which allows translation to continue through a stop
codon . Suppressor tRNA tend to be found in cells in
which a mutation has introduced a stop codon.
EUKARYOTIC TRANSLATION

 Coupled Transcription and Translation in

Eukaryotes

- Until recently, the dogma of eukaryotic translation

was that it was physically separated from transcription.
Transcription occurred in the nucleus, and mRNA was
then exported to the cytosol for translation. Although
this system is accepted as the normal process, recent
evidence has shown that the nucleus has all of the
components (mRNA, ribosomes, protein factors)
necessary for translation.
Posttranslational Modification
of Proteins
Posttranslational Modification of
Proteins

- Newly synthesized polypeptides are frequently

processed before they reach the form in which they
have biological activity. Proteins destined for export to
specifi c parts of the cell or from the cell have leader
sequences at their N-terminal ends. These leader
sequences, which direct the proteins to their proper
destination, are recognized and removed by specifi c
proteases associated with the endoplasmic reticulum.
The fi nished protein then enters the Golgi apparatus,
which directs it to its fi nal destination.
Posttranslational Modification of
Proteins
Some examples of posttranslational modification of
proteins
 Posttranslational Modification of
Proteins

Once modified, do proteins always

have the correct three-
dimensional structure?
 Posttranslational Modification of
Proteins

 In theory, the primary structure of the protein

determines its three-dimensional structure. However, in
reality proteins often need the help of a chaperone to
arrive at the correct structure.
Protein Degradation
 Protein Degradation

- One of the most often overlooked controls of gene

expression occurs at the level of the degradation of
proteins. Proteins are in a dynamic state in which
they are turned over often.
 Protein Degradation

How does the cell know which

proteins to degrade?
 Protein Degradation

• Proteins are degraded in subcellular organelles, such

as lysosomes, or in macromolecular structures called
proteasomes.
• Many proteins are targeted for destruction by being
bound to a protein called ubiquitin. The nature of the
amino acid sequence at the N-terminus is often very
important to control the timing of the destruction of a
protein.
• Damaged proteins are degraded very quickly.
 Protein Degradation

Diagram of the ubiquitin–

proteasome degradation
pathway
 Protein Degradation

Degradation of
proteins with acidic
N-termini
THANK YOU!!!!!

SUBMITTED BY: GROUP 10

DAGAMI, RICHEL
GALAN, RONALIZA
LERIOS, MARIANNE ANGELI
MARTIJA, CHERRY KAY
SEROJIHOS, JOY