TRANSLATION IN PROKARYOTES

AND GENETIC CODE…

SUBMITTED TO: Dr. P P BALGIR

SUBMITTED BY: VANITA
CLASS : MSc. BIOTECHNOLOGY (I)
ROLL No.: 18011010
CONTENTS:
Flow of genetic material
Introduction
Genetic code
Tools for translation:
-Ribosome
-tRNA
Initiation in prokaryotes
Elongation in prokaryotes
Termination in eukaryotes
FLOW OF GENETIC MATERIAL:

• The central dogma of molecular biology describes the two step

process , transcription and translation , by which the information in
genes flows into proteins :

DNA RNA PROTEIN

INTRODUCTION OF TRANSLATION:
• It is a well conserved process among prokaryotes and eukaryotes.
• Translation: the biosynthesis of a protein or a polypeptide inside a
living cell.
• It is a universal process.
• Protein synthesis is the final stage of gene expression.
• It occurs in cytoplasm where ribosome are located.
• The genetic message transcribed to mRNA is translated into protein
by a complex cellular machinery. Additional processing and assembly
often required to modify the proteins
• In process of translation the language of nucleotides sequence on
mRNA is translated into the language of amino acid sequence.
• In translation, messenger RNA is decoded to produce a specific
polypeptide.
• This uses mRNA sequence as a template to guide the synthesis of a
chain of amino acid that form protein.
• Many types of transcribed RNA, such tRNA, rRNA, snRNA are not
necessarily translated to amino acid sequence.
• The process of translation requires a genetic code , through which the
information contained in nucleic acid sequence is expressed to
produce a specific sequence of amino acids.
GENETIC CODE:
• The letters A, G , T , C corresponds to the nucleotide found in DNA.
They are organized into codons.
• The collection of codons is called genetic code.
• The genetic code is the system of nucleotide sequences of mRNA that
determines the sequence of amino acids in protein.
• Codons are a group of three adjacent bases that specify the amino
acids of protein.
• Since there are 4 bases and 3 positions in each codon, there are
4×4×4 =64 possible codons .
• 64 codons but only 20 amino acids , therefore most have more than 1
codon.
• 3 of the 64 codons are used as stop signals(UAA, UAG,UGA)
• 1 codon is used as a start codon(AUG)
CHARACTERISTICS OF GENETIC CODE:
• Triplet codons
• Number of codons
• Stop or termination or nonsense codons
• Degenerate
• Unambiguous
• Universal
• Non- overlapping and nonpunctuation
1. INITIATOR CODON:
• AUG is the initiator codon in majority of proteins.
• In a few cases GUG may be the initiator codon.
• Methionine is the only amino acid specified by just one codon ,AUG.

2. SPECIFICITY:
• Genetic code is specific (unambiguous).
• A specific codon always codes for the same amino acid.
e.g. UUU codes for phenyl alanine, it can not code for any other amino
acid.
3. UNIVERSAL: In all living organism genetic code is the same.
• There are some exceptions for this universality
e.g. : AGA and AGG code for arginine in cytoplasm but in mitochondria
they are termination codons.

4. DEGENERATE:
• Genetic code is redundant, also called degenerate.
• Although each codon corresponds to a single amino acid but a single
amino acid can have multiple codons.
• Except Tryptophan and Methionine each amino acid has multiple
codons.
5. NON OVERLAPPING AND NON PUNCTUATED:
• All codons are independent sets of 3 bases.
• There is no overlapping.
• Codon is read from a fixed starting point as a continuous sequence of
bases.
• The starting point is extremely important and this is called reading
frame.
6. NON SENSE CODONS:
• There are 3 codons (UAA,UAG,UGA) out of 64 in genetic code which
do not encode for any amino acid.
• Also called termination codons or stop codons.
• The ribosome pauses and falls off the mRNA.
WOBBLING PHENOMENON:
• The rules of base pairing are relaxed at the third position , so that a
base can pair with more than one complementary base.
• Some tRNA anticodons have inosine at the third position which can
pair with U,C, or A.
• The reduced specificity between the
third base of the codon and the
complementary nucleotide in anticodon
is responsible for wobbling.
TOOLS OF TRANSLATION:
• Ribosomes
• tRNA
• mRNA
• Mg 2+
• Amino acids
• Initiation ,elongation , termination factors
• Amino acyl tRNA synthetases ( I and II)
transfer RNA:
• 3’ end of tRNA : binding
site for amino acids.
• Anticodon loop at
opposite end: interacts
with complementary
codon on mRNA.
RIBOSOMES:
• Ribosomes are the macromolecular complex that directs the
synthesis of proteins.
• These are the sites of protein synthesis having
-30% -40% protein
-60%-70% RNA
• Each ribosome having 2 ribosomal subunit- larger and smaller.
• In prokaryotes ribosome are 70s and in eukaryotes ribosomes are
80s.( s stands for svedbergs a unit to measure how fast molecule
move in a centrifuge.)
• Ribosomes has three tRNA binding site:
A site- binding site for first aminocylated tRNA.
P site- binding site for the peptidyl tRNA.
E site- binding site for the uncharged tRNA.
• These sites are present at the interface
between the small and the larger subunit
of ribosome.
TRANSLATION IN PROKARYOTES:
• Translation proceeds in three steps:
1. Initiation
2. Elongation
3. Termination

Before these steps activation of amino acid is necessary which is

required for translation to proceed.
ACTIVATION OF AMINO ACID :

• It involves 2 steps:
1. Adenylation of amino acid
2. Transfer of adenylated
amino acid to the tRNA.

The enzyme involved is

Aminoacyl-tRNA synthetase.
INITIATION:
• Prokaryotes initiation require the large and small subunits , the mRNA, the
initiator tRNA, three initiation factors(IF -1, IF-2, IF-3) and GTP.
• IF-3 binds to the free 30s subunit , this help to prevent larger subunit
binding to it without mRNA and forming an inactive ribosome.
• IF-2 complexed with GTP
• IF-1 binds to small subunit. It will assist the charged initiator tRNA to bind.
• It involves 2 steps:
1. Formation of 30s complex
2. Formation of 70s initiation complex
FORMATION OF 30s COMPLEX:
The 30s subunit attached to a mRNA molecule making use of the
ribosomal binding site on mRNA(shine Dalgarno sequence that is
UCCUC: 16 s rRNA).
Interaction between theses complementary sequence enhances the
attachment of the 30s subunit to the AUG initiator codon.
The initiator tRNA can then bind to the complex by base pairing of its
anticodon with AUG codon on mRNA.
Now, IF3 can be released , as its role in keeping the subunits apart are
complete.
This complex is called 30s initiation complex.
FORMATION OF 70s COMPLEX:
The 50s subunit can now bind , which displace IF1 and IF2, and the
GTP is hydrolysed in this energy consuming step.
This complex is called 70s initiation complex.
ELONGATION:
• With the formation of 70s initiation complex the elongation cycle can
begin.
• It involves three elongation factors EF-Tu, EF-Ts and EF-G, GTP,
charged tRNA and the 70s initiation complex.
• Elongation is divided into 3 steps:
1. Amino –acyl tRNA delivery
2. Peptide bond formation
3. Translocation
1. AMINO- ACYL tRNA DELIVERY:
• EF-TU is required to deliver the amino acyl tRNA to A site and energy
is consumed in this step by hydrolysis of GTP.
• The released EF-Tu GDP complex is regenerated with the help of EF-
TS.
• In the EF-Tu EF-Ts exchange cycle EF-Ts displaces the GDP and replace
itself by GTP.
• The resultant EF-Tu . GTP complex is now available to bind another
amino acyl tRNA and deliver it to ribosome.
• All amino acyl tRNA can form this complex with EF-Tu except the
initiator tRNA.
2. PEPTIDE BOND FORMATION:
• After aminoacyl – tRNA delivery , the A and P sites are both occupied
and the two amino acids that are to be joined are close to each other.
• The peptidyl transferase activity of the 50s subunit can now form a
peptide bond between the two amino acids.
3. TRANSLOCATION:
• A complex of EF-G( translocase) and GTP binds to the ribosome and is
an energy consuming step, the discharged tRNA is ejected from the P
site, the peptidyl- tRNA is ejected from the P site , the peptidyl -tRNA
is moved from A site to P site.
• The mRNA moves by one codon relative to one codon to the
ribosome.
• GDP and EF-G are released . A new codon is now present in the
vacant site.
 AMINO-ACYL tRNA
DELIVERY

TRANSLOCATION
PEPTIDE BOND
FORMATION
TERMINATION:
• Termination of translation happens when A site of the ribosome faces
a stop codon (UUA, UGA, UGA)
• Then no tRNA can recognise it, but a releasing factor can recognize
the stop codons and cause the release of polypeptide chain.
• In prokaryotes once a stop codon occupies the A site , three
termination or release factor (RF1, RF2,RF3) contribute to the
hydrolysis of peptidyl –tRNA bond.
• Release the free polypeptide and last uncharged tRNA from P site.
• The dissociation of the 70s into 30s and 50s subunit.
Functions:
• RF1 binds A site and release the polypeptide and uncharged tRNA.
• RF2 releases the RF1 from A site and release itself as well from
translation binding site .
• RF3 function unknown.
• Another factor called ribosomal releasing factor causes the
dissociation of 70s complex.
REFERENCES:
• GENES V : BENJAMIN LEWIN
• LIFESCIENCES
• www.biologydiscussion.com
THANKYOU…