TRANSLATION

By: Poonam Sharma M.Sc.Biotech.4th sem.

TRANSLATION: Synthesis of protein

Translation - genetic information carried by the DNA is decoded, using an RNA intermediate, into proteins. DNA-RNA- Protein Proteins - structural constituent of a cell. Bacterial translation. Eukaryotic translation. (i) Protein Synthetic machinery. (ii) Mechanism of protein synthesis.
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(i) Protein synthetic machinery: Amino acids DNA as specificity control RNAs Ribosomes Enzymes (ii) Mechanism of protein synthesis: Activation of amino acids Charging of tRNA Activation of ribosomes Assembly of amino acids
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Activation of amino acids and charging of tRNA

Amino acid

ATP

PPi

Specific activating enzyme

Amino acid - AMP

Specific tRNA
Charging enzyme

AMP

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Amino acid

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Charged tRNA

Activation of Ribosomes
activation of ribosomes - by mRNA chain.

The small ribosomal subunits joins by first codon through base pairing with appropriate sequence on rRNA. And the combination is called initiation complex. larger ribosomal subunits joins the small subunits forming active ribosome. Requires Mg2+ ion
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ASSEMBLY OF AMINO ACIDS

better known in bacteria than eukaryotes.
The focus here will be on structural aspects, and on protein factors involved in: initiation. elongation, & termination of amino acid chain.

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Initiation Of Polypeptide Chain:

Initiation of protein synthesis in E. coli requires initiation factors IF-1, IF-2, & IF-3. IF-1 assists binding of IF-3 to the 30S ribosomal subunit IF-2 is a small GTP-binding protein

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Initiation of translation in prokaryotes

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Elongation Of Polypeptide Chain:

Elongation - 70s ribosome. Elongation does not accept the formylmethionine tRNA, but accepts methionine tRNA in the presence of elongation factors. The ribosome has 2 sites for tRNAs, called P and A

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Elongation requires participation of elongation factors EF-Tu (also called EF1A) EF-Ts (EF1B) EF-G (EF2) EF-Tu & EF-G are small GTP-binding proteins.

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Factor EF-Tu helps in aminoacyl tRNA entry

Factor EF-G helps in translocation and enzyme is Peptidyl transferase

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CHAIN TERMINATION
Three codons are called stop codons. They code for no amino acid, and all protein-coding regions end in a stop codon.

Chain termination requires release factors RF-1, RF-2, & RF-3.

RF-3 is a small GTP-binding protein

RF-1 & RF-2 recognize & bind to STOP codons. One or the other binds when a stop codon is reached.
RF-3-GTP facilitates binding of RF-1 or RF-2 to the ribosome.
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RFI, RF2 and RF3 are release factors

rRF1and RF2 bind to the stop codon

RF3 facilitates the binding of RF1 and RF2 to ribosome

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Eukaryotic Translation
Translation of mRNA is highly regulated in multi-cellular eukaryotic organisms, whereas in prokaryotes regulation occurs mainly at the level of transcription. There is global regulation of protein synthesis. E.g., protein synthesis may be regulated in relation to the cell cycle or in response to cellular stresses such as starvation or accumulation of unfolded proteins in the endoplasmic reticulum. Mechanisms include regulation by signal-activated phosphorylation or dephosphorylation of initiation and elongation factors.
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Translation of particular mRNAs - inhibited by small single-stranded microRNA - 20-22 nucleotides long. MicroRNAs bind via base-pairing to 3' un-translated regions of mRNA along with a protein complex RISC (RNA-induced silencing complex), inhibiting translation and in some cases promoting mRNA degradation. Protein factors that mediate & control translation are more numerous in eukaryotes than in prokaryotes.

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Eukaryotic factors are designated with the prefix "e".

Some factors are highly conserved across kingdoms. E.g., the eukaryotic elongation factor eEF1A is structurally and functionally similar to the prokaryotic EF-TU (EF1A). In contrast, eEF1B, the eukaryotic equivalent of the GEF EF-Ts, is relatively complex, having multiple subunits subject to regulatory phosphorylation. eEF2:translocation factor which is like EF-G

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Initiation of protein synthesis is much more complex in eukaryotes, & requires a large number of protein factors.
Some eukaryotic initiation factors (e.g., eIF3 & eIF4G) After the initiation complex assembles, it translocates along the mRNA in a process called scanning, until the initiation codon is reached. Scanning is facilitated by eukaryotic initiation factor eIF4A, which functions as an ATP-dependent helicase to unwind mRNA secondary structure while releasing bound proteins.

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Initiation Of Translation In Eukaryotes

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A short sequence of bases adjacent to the AUG initiation codon may aid in recognition of the start site.
After the initiation codon is recognized, there is hydrolysis of GTP and release of initiation factors, as the large ribosomal subunit joins the complex and elongation commences. Some eukaryotic mRNAs have what is called an internal ribosome entry site (IRES), far from the 5' capped end, at which initiation may occur without the scanning process.

Eukaryotes have single release factor eRF

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