PROTEIN SYNTHESIS

Francis Ian L. Salaver, RMT

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The Genetic Code

Use the code by reading from the center to the outside Example: AUG codes for Methionine

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Name the Amino Acids

GGG? UCA? CAU? GCA? AAA?
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Transfer RNA (tRNA)

Clover-leaf shape Single stranded molecule with attachment site at one end for an amino acid Opposite end has three nucleotide bases called the anticodon
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Transfer RNA
amino acid attachment site

U A C
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anticodon

Aminoacyl-tRNA synthetases

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Two-stage reaction
In the first stage, the amino acid is activated with ATP. In the second stage of the reaction, the amino acid is attached with a highenergy bond to the 2 or 3 carbon of the ribose sugar at the 3 end of the transfer RNA

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Codons and Anticodons

The 3 bases of an anticodon are complementary to the 3 bases of a codon Example: Codon ACU Anticodon UGA
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UGA

ACU

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Transcription and Translation

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Pathway to Making a Protein

DNA
mRNA tRNA (ribosomes) Protein

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Protein Synthesis
The production or
synthesis of polypeptide chains (proteins) Two phases: Transcription & Translation mRNA must be processed before it leaves the nucleus of eukaryotic cells
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DNA RNA Protein

DNA
Transcription Nuclear membrane

Eukaryotic Cell

Pre-mRNA

RNA Processing

mRNA

Ribosome
Translation

Protein
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Translation
Translation is the process of decoding the mRNA into a polypeptide chain Ribosomes read mRNA three bases or 1 codon at a time and construct the proteins
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Transcription

Translation

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Ribosomes
Made of a large and small subunit Composed of rRNA (40%) and proteins (60%) Have two sites for tRNA attachment --- P and A
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Translation has 3 phases

1. Initiation 2. Elongation 3. Termination

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Initiation of Translation
In prokaryotes, ribosomes bind to specific translation initiation sites. There can be several different initiation sites on a messenger RNA: a prokaryotic mRNA can code for several different proteins. Translation begins at an AUG codon, or sometimes a GUG. The modified amino acid Nformyl methionine is always the first amino acid of the new polypeptide.

Initiation of Translation
In eukaryotes, ribosomes bind to the 5 cap, then move down the mRNA until they reach the first AUG, the codon for methionine. Translation starts from this.
Note that translation does not start at the first base of the mRNA. There is an untranslated region at the beginning of the mRNA, the 5 untranslated region (5 UTR).

Step 1- Initiation
mRNA transcript start codon AUG attaches to the small ribosomal subunit Small subunit attaches to large ribosomal subunit
mRNA transcript
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More Initiation
The initiation process involves first joining the mRNA, the initiator methionine-tRNA, and the small ribosomal subunit. Several initiation factors-additional proteins--are also involved. The large ribosomal subunit then joins the complex.

An initiation codon marks the start of an mRNA message

AUG = methionine
Start of genetic message

End

Figure 10.13A

 mRNA, a specific tRNA, and the ribosome subunits assemble during initiation

Initiator tRNA P site

Large ribosomal subunit A site

Start codon mRNA

Small ribosomal subunit

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Figure 10.13B

Elongation
The mRNA moves a codon at a time relative to the ribosome
A tRNA pairs with each codon, adding an amino acid to the growing polypeptide A STOP codon causes the mRNA-ribosome complex to fall apart

Elongation

Amino acid Polypeptide P site mRNA 1 Codon recognition A site

Anticodon

mRNA movement

Stop codon
New peptide bond 2 Peptide bond formation

Translocation

Figure 10.14

Ribosomes
Large subunit

P Site

A Site
mRNA

Small subunit

A U G

C U A C U U C G
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Initiation
aa1 aa2

2-tRNA 1-tRNA

anticodon
hydrogen bonds

U A C A U G codon

G A U C U A C U U C G A
mRNA
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Elongation
peptide bond
aa1 aa2 aa3

3-tRNA 1-tRNA 2-tRNA

G A A

anticodon
hydrogen bonds

U A C A U G codon

G A U C U A C U U C G A
mRNA
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aa1

peptide bond

aa3
aa2

1-tRNA

U A C
(leaves) 2-tRNA

3-tRNA

G A A

A U G

G A U C U A C U U C G A
mRNA
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Ribosomes move over one codon

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aa1

peptide bonds

aa4
aa2 aa3

4-tRNA
2-tRNA 3-tRNA

G C U

A U G

G A U G A A C U A C U U C G A A C U
mRNA
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aa1

peptide bonds aa2 aa3

aa4

2-tRNA

G A U
(leaves) 3-tRNA

4-tRNA

G C U

A U G

G A A C U A C U U C G A A C U
mRNA
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Ribosomes move over one codon

aa1

peptide bonds aa2 aa3 aa4

aa5

5-tRNA

U G A
3-tRNA 4-tRNA

G A A G C U G C U A C U U C G A A C U
mRNA
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aa1

peptide bonds

aa5

aa2
aa3 aa4

5-tRNA

3-tRNA

U G A
4-tRNA

G A A

G C U G C U A C U U C G A A C U
mRNA

Ribosomes move over one codon

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aa4

aa5
aa199
aa200

Termination

aa3 primary structure aa2 of a protein aa1

200-tRNA

terminator or stop codon

A C U
mRNA

C A U G U U U A G
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Termination
Three codons are called stop codons. They code for no amino acid, and all protein-coding regions end in a stop codon. When the ribosome reaches a stop codon, there is no tRNA that binds to it. Instead, proteins called release factors bind, and cause the ribosome, the mRNA, and the new polypeptide to separate. The new polypeptide is completed.

End Product The Protein!

The end products of protein synthesis is a primary structure of a protein A sequence of amino acid bonded together by peptide bonds
aa2 aa1 aa3 aa4 aa5
aa199 aa200 38

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