PROTEIN SYNTHESIS (TRANSLATION)

C-4 Tutorial Group

Codon
It is now known that each codon consists of a sequence of three nucleotides a triplet code

Start codon AUG methionin

Stop codons UAG, UGA, UAA

The Characteristics of Codon

Degenerate multiple codons must decode the same amino acid Unambiguous given a specific codon, only a single amino acid is indicated Non-overlapping the process of protein synthesis does not involve any overlap of codons

The Characteristics of Codon

Not punctuated the message is read in a continuing sequence of nucleotide triplets until a translation stop codon is reached Universal The frequency of use of each amino acid codon varies considerably between species and among different tissues within a species

Components of Translation
Amino acid all amino acid must be present at the time of protein synthesis tRNA attach to specific amino acid, contains anticodon Aminoacyl-tRNA synthase attach the amino acids to their corresponding tRNAs

Components of Translation
mRNA as a template for the synthesis of the polypeptide chain Ribosom
rRNA structure of base-pairing Ribosomal protein interact with other component of translation system A-binding site aminoacyl tRNA P-binding site peptidyl tRNA E-site exit of the ribosome

Components of Translation
Protein factors as catalytic factors / enzymes ATP & GTP source of energy

Steps in Protein Synthesis

Initiation Elongation Termination

Postranslation Modification

Initiation
The binding of GTP by eIF-2 binary complex Binary complex then binds to met-tRNA ternary complex Ternary complex binds to the 40S ribosomal subunit the 43S preinitiation complex The 43S preinitiation complex stabilized by association with eIF-3 and eIF-1A.

Initiation
The association of mRNA with the 43S preinitiation complex + ATP hydrolysis 48S initiation complex 60S ribosomal subunit + 48S initiation complex + hydrolysis of the GTP bound to eIF-2 by eIF-5 = release of the initiation factors bound to the 48S initiation complex 40S and 60S subunits to form the 80S ribosome At this point, the met-tRNAi is on the P site of the ribosome, ready for the elongation cycle to commence.

Elongation
Binding of aminoacyl-tRNA to the A site aminoacyl-tRNA carries out a nucleophilic attack on the esterified carboxyl group of the peptidyl-tRNA occupying the P site by peptidyltransferase

Elongation
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 energy requirements for the formation of one peptide bond = hydrolysis of two ATP molecules to ADP and of two GTP molecules to GDP = hydrolysis of four highenergy phosphate bonds.

Termination
The elongation will stop terminating codon of mRNA if stop codons (UAA, UAG, UGA) appears in the A site The releasing complex (Releasing factor RF1 + releasing factor RF3 + GTP) hydrolysis peptide bonds and release protein and dissociate the 80S into 40S and 60S

The Regulation of eIF-4E Controls the Rate of Initiation

The 4F complex is particularly important in controlling the rate of protein translation. 4F is a complex consisting of 4E, which binds to the m7G cap structure at the 5 end of the mRNA, and 4G, which serves as a scaffolding protein. In addition to binding 4E, 4G binds to eIF-3, which links the complex to the 40S ribosomal subunit. It also binds 4A and 4B, the ATPase-helicase complex that helps unwind the RNA. 4E is responsible for recognition of the mRNA cap structure, which is a rate-limiting step in translation.

REGULATION ON TRANSLATIONAL STAGE

In eukaryotes, eukaryotic initiation factor 2 (eIF2) is the center of regulation mechanism as eIF2 is needed in initiation stage of protein synthesis. eIF2 can be inhibited by phosphorylation.

REGULATION ON POSTTRANSLATIONAL STAGE

Post-translational modification is defined as protein structure alteration after being released by ribosome. Modification takes place in endoplasmic reticulum and Golgi apparatus. This modification can be classified into two groups:
Chemical modification. Protein folding.

Posttranslational Modification
The result of translation proproteins require modifications before attaining biologic activity Kind of posttranslational removal amino acid residues by aminopeptidases acetylation phosphorylation methylation ubiquitinylation glycosylation