1

TRANSLATION
The central dogma of life

DNA

transcription

mRNA

translation

protein

THE GENE

The code has 4 letters- A,C,G & T (4 bases)

The code is composed of triplet codes/codons. eg - AGC, CTC, TGG
Each codon code for one amino acid.
3
There are 64 diferent codones. ( 4 =64 )

But only 61 of them code for amino acids.

The other 3 do not- stop codons
A series of codons in DNA that coding for a protein is a GENE.

Characteristics of the genetic code

a. Unambiguous
The code occur as codons. Each codon code for only one amino acid.
b. Degenerate
But a amino acid may have more than one codon. eg;- Ala - 4 codons
c. Non overlapping
Code is read,
1. From a fixed starting point. (AUG)
2. 3 bases at a time
3. No punctuation
d. Universal (almost)
The code is almost the same for all the living organisms.
Why almost?
Because there are minor variations
1. The amino acid for start codon (AUG)
In eukaryots- Meth
In prokariotes- fMeth
2. Variations in mitochondrial genome, chloroplast & some ciliated protozoa. Eg;AGA is an stop codon in mitochondrial genome
Repeat campaign A/L 2007

2
Translation
Converting the coded message in the mRNA the amino acid sequence of the protein
Translation requires-mRNA, tRNA, ribosomes, initiation factors(proteins)

tRNA
e. 3 end carries the amino acid.
f. base pairing is antiparallel.(3rd base of codon
pair with 1st base of anticodon)
g. wobble hypothesis
pairing of 3rd base in codon with 1st base of anticodon
not strictly by Watson-Crick rule.
h. Therefore more than one tRNA can bring
one amino acid
eg;- anticodon- (3) XYU (5)
codon
-(3) YXA (5)
(3) YXG (5)
For one amino acid

Formation of amino acyl tRNA

Step 1 :Step 2 :-

AA + ATP Aminoacyl AMP+ PPi

Aminoacyl AMP+tRNA AminoacyltRNA+ AMP

Both activities are catalized by Aminoacyl tRNA synthetase

It has a proof reading activity to ensure that the correct amino acid is bound to
the tRNA.
3. 1st check (represents the 1st step)
If wrong amino acyl AMP is bound to the enzyme,
it is hydrolyzed to amino acid and AMP
4. 2nd check (represents the 2nd step)
If wrong amino acid is bound to tRNA,
The ester linkage of amino acyl tRNA is hydrolyzed.
There are 20 aminoacyl tRNA synthetases, each specific for one amino acid.
But there are 60 different tRNAs. The tRNAs that can bind the same amino acid
are recognized by the same enzyme.
Repeat campaign A/L 2007

3
Specific nucleotides on tRNAs are involved in this recognition by the enzyme.this
is called second genetic cord

Aminoacyl tRNA

Ribosomes :- eukaryotic-

80 S 40 S+60 S

prokaryotic-

70 S 30 S +50 S

amino acid

Ester bond

Prokaryotic DNA translation.

Three steps :-

initiation elongation termination

Initiation
1.

Ribosomes separate(IF1 & IF3 help and prevent recombination)

70 S 30 S +50 S

2. 30S subunit join mRNA near 5 end, the P site opposite the start/initiation
codon(AUG). (Shine- Delgano sequence just upstream of AUG help in oppositioning)
3. fMet-tRNA bind to AUG forming 30S initiation complex. ( GTP bound IF2 help this)
4. 50S subunit bind to 30S initiation complex, GTP hydrolyzed, 70S initiation complex
formed.

1
A 2
5 U
P
U3A
X
A
C0

3
X
G X X
S

30S initiation complex

1
5
0
U
A
S32 3
PA
A
U0X X
C
GSX X

70S initiation complex

12

AG
23
5U
A
A
UA
XX
P
A
C
GXX

Repeat campaign A/L 2007

4
Elongation
1. Elongation factor Tu(EF Tu), GTP & aminoacyl-tRNA, form a complex and and bind to
ribosome at A site.
2. If anticodon does not match, complex dissociate.
3. If matches, GTP is hydrolyzed EF-Tu-GDP complex leave the ribosome.
4. Peptide bond formation
12
23S RNA of 50S ribosome has peptidyl-transferase
U
activity.
AG
23
A
A
Ester bond between fMet & tRNA is broken , fMet is
UXX
C
transferred to A site, peptide bond is formed between 1 st &
GA
XX
nd
2 amino acid at A site.
5. Translocation of ribosome
GTP binding EF-G (a translocase) hydrolyse GTP and provide
energy.
Ribosome changes its shape and moves to the next triplet
1 23
cord, in 3 direction.
Dipeptide now occupy P site, tRNA at AUG codon leaves the U
A A G
3
2 C
ribosome.
A
G
C
6. New amino acyl tRNA arrives at empty A site.
UX X
A X
7. The process continues till the stop codon is reached.
GXA

5 PA 3

3
PA

Termination

1. When A site occupy a termination codon(UAA,UGA,UAG) a Releas Factor(RF) binds to

it(prokaryotes- RF1, RF2,RF3 , eukaryotes- eRF)
2. Protein tRNA bond hydrolyzed releasing the protein & tRNA.
3. Ribosome dissociate to start a new cycle.

6
45 78
3

6
45 718
3

R
U 3
8F
5 7G
A
XA
XPA
A

R
G
F
5A 7 8 U
XA
A X PA
XXG

XXG

Polyribosome/ Polysome- several ribosomes attached to one mRNA.

Can be free in cytosol or attached to ER (RER)

Antibiotics that inhibit ribosomal protein synthesis

Drug

subunit

Target organism

effect
Repeat campaign A/L 2007

5
Streptomycin
Tetracycline
Chloramphenicol
Erythromycin
Puromycin

Cycloheximide

30S
30S
50S
50S
50S

prokaryotes
prokaryotes
prokaryotes
prokaryotes
prokaryotes &

Stop initiation & misread mRNA

Stop aminoacyl tRNA binding
Inhibit peptidyl transferase
Inhibit translocation
Structural analogue of tyrosinyl-tRNA

60S

eukaryotes

incorporate at c-treminal & stop

eukaryotes

elongation
Inhibit peptidyl transferase

60S

Effect of diphtheria toxin on protein synthesis

a.
b.
c.
d.
e.
f.
g.
h.

Organism- Corynebacterium diphtheria, a bacterium

The toxin bind to receptors on mucosal cell surface & is proteolytically cleaved.
one fragment the cell and
inactivate elongation factor 2 (EF2)
It is equivalent to prokaryotic EF-G
EF2 is required for the GTP driven translocation of ribosome along mRNA.
Stop protein synthesis & kills the cell.
Act as an enzyme & small amount is sufficient for a great damage.
(any 6 x 5= 30)

Signal sequence
a. 20-25 AAs long portion at the amino terminal end of export proteins.
b. Contain high proportion of hydrophobic AAs
c. helps export proteins from inside the cell to the out side.

Post translational modifications

Changes that a translated protein undergo to form the final Biologically Active Protein
1. Amino/carboxyl terminal modifications
a. Formyl group of amino terminal Met frequently removed.
b. Acetylation of amino terminal residue in many eukaryotic proteins.
2. Removal of Signal sequence. Eg:- ALA synthase
3. Formation of disulfide bonds between Cys: residues
4. Proteolytic cleavage of parts of polypeptide chain.
a. Removal of the c-peptide from insulin.
b. Removal of the hexapeptide from the amino terminal converts trypsinogen to
active trypsin.
c. Removal of the 44 AAs from the amino terminal converts pepsinogen to active
pepsin.
5. Covalent adition of prosthetic groups.
a. addition of biotin to Acetyl CoA Carboxylase
b. addition of heme group to Cytochrome C
6. Modifications of individual AAs
Repeat campaign A/L 2007

6
a. Phosphorylation of OH groups of Ser, Thr & Tyr
I.
Milk protein casein has many Phosphoserines.
Net negativity of the protein increases.
Help bind Ca2+
II.
Uncontrolled Phosphorylation of certain Tyr residues of some proteins
induce cancer.
Eg:- erbB oncogene product- erbB protein (a membrane receptor)
continuously phosphorylate proteins & lead to adenocarcinomas
b. Hydroxylation
Proline & Lysine residues of procollagen are hydroxylated (by Prolyl Hydroxylase,
Lysyl Hydroxylase) during the maturation process.
This process requires Ascorbic acid.
A deficiency leads to poor hydroxylation of collagen.
Poor cross linking between fibrils leads to Scurvy

7. Glycosylation
a. enzymetic glycosylation at
Amide nitrogen of Asparagine
Hydroxyl groups of Ser or Thr
b. nonenzymetic Glycosylation of Hb
occur at N-terminal NH2 groups.
Glycosylated Hb concentration reflects glycaemic status during entire life
span of a RBC .(accurately monitor 6th to 8th week of time)
Useful to check glycaemic control of diabetis.

8. I- cell disease
A carbohydrate moiety- an oligosaccharide containing mannose 6-PO4 is
attached to the lysosomal proteins.
mannose 6-PO4 is bound by a specific Glycosyltransferase.

mannose 6-PO4 acts as a Targeting signal that is identified by receptor that

target the protein into lysosomes.
Lack of the Glycosyltransferase lead to glycoproteins without the
mannose 6-PO4 residue.
Targeting process of enzymes to lysosome is deficient.
The enzymes are secreted out of the cell.
Lysosomes accumulate partly digested materials that manifest as
Inclusion bodies.
Any 6 x 5=30
Repeat campaign A/L 2007

Repeat campaign A/L 2007