TRANSLATION

Components for translation

mRNA transmits info encoded in DNA
Eukaryotic
A 5UTR precedes the translation initiation
signal
Usually the 1st AUG sequence (initiation codon)
Prokaryotic
No capped
Polycistronic
More than 1 initiation codon
Ribosome positioning sequence at 10
nucleotides upstream of the AUG codon
UTR follows termination signal
tRNA is a bilingual translator molecule
Clover-leaf = secondary structure
L-shaped = tertiary structure
Codons in mRNA are 3-letter words
1:1 correspondence of bases to AA = permit
mRNA to encode only 4 diff. AA
2:1 correspondence of bases to AA = 16 AA
Multiple codons for a single amino AA
degeneracy
Start signal (AUG) = methionine signifies the
initial, amino terminal residue
UGA = termination (usual)
Mitochondrial termination = AGA, AGG
Stop signals = UAG, UAA, UGA
UGA codon = additional function of coding for
SELENOCYSTEINE
Seleocysteine-tRNA is synthesized from a
specific seryl-tRNA, a sec-inserion sequence, a
defined stem-loop structure in mRNA
o eEFSec & SBP2 (selenocysteine
insertion binding protein required for
its synthesis
Codon-Anticodon interactions permit reading of mRNA
anti-parallel
Codons are read in a sequenctial, nonoverlapping reading frame
Anticodon & AA-acceptor sites are at opposite
ends of L-shaped tRNA
o Bridges the gap between nucleotide
sequence of mRNA & site of protein
assembly on ribosome
Frequent anticodon nucleotide is inosinic acid
(I), hypoxanthine, which base pairs with U, C, A
o Wobble hypothesis = permits less
stringent base pairing between the 1st
position of anticodon and the
degenerate (3rd) position of codon
3codon
base
A
C
G

5 anticodon base
possible
U, I
G, I
C, U

A, G, I

Aminoacylation of tRNA activates AA for protein

synthesis
Two step process catalyze by aminoacyl-tRNA
synthetases (each is specific for single AA)
Aminoacyl-ester linkage is lower in energy
compared to aminoacyl-adenylate
Correct selection of both tRNA and AA by the
synthetase is therefore central to fidelity in
translation
Some AA are recognized by bulk (Try) o lack of
bulk (Gly) or by charges
Additional proofreading step increases
discrimination by many (not all) aminoacyl
synthethases
Each synthetase must also correctly recognize
one to several tRNA species that can carry the
same AA, while rejecting incorrect ones.
Ribosomes
Complex RNPs that is made up of 2 dissimilar
subunits, each contains RNA & many proteins
Large subunit sites of peptide bond formation
(peptidyl transferase) ad of elongation factor
binding (translocation domain) opposite site of
central protuberance
Stalk projects to right
Small subunit platform protrudes toward the
reader.
mRNA & tRNA interact in the decoding site, in a
cleft between platform & subunit body
the site of peptide bond formation is distant
from exit site and the growing peptide passes
through groove in ribosome to reach the exit
aminoacyl-tRNA is bound by the small subunit
an is oriented with the acceptor end near
peptidyl transferase
while translocation domain is near the
decoding site
tRNA is directional & collinear with mRNA
also 5-3
INITIATION OF PROTEIN SYNTHESIS
Requirements: (should be in proper orientation)
1) Small (40S) ribosomal subunit
2) mRNA
3) tRNA complex of amino-terminal
AA
4) followed by association of large
(60S) subunit now a completed
initiation complex
STEPS:
1) eIF2 binds to GTP & Met-tRNA
(initiator tRNA) forming TERNARY
COMPLEX

no other aminoacyl-tRNA
can replace initiation
specific Met-tRNA in this
step

8) GTP is hydrolyzed eIF5B-GTP is

released
9) Complete initiation complex is an
80S ribosome with mRNA &
initiator tRNA correctly positioned
to begin translation

2)

3)

4)
5)
6)

7)

Prokaryotes: fMet-tRNA
(formylated); recognized by
IF2
eIF3 binds to small subunit (40S)
surface blocking subunit
association with 60S

eIF3 = anti-association
factor

eIF6 = binds 60S subunit

(+) COMPLEX

eIF2-GTP

Met-tRNA

eIF3-40S
PRE-INITIATION COMPLEX

mRNA-eIF4F (cap-binding
complex)

eIF4A = RNA helicase;

unwinds secondary
structure in untranslated
leader sequence of mRNA

PAB (Poly A binding

protein) = loops the 3 end
of mRNA near to 5 cap
mRNA is scanned in 5 3 until
AUG is found
GTP is hydrolyzed via eIF5 eIF2GDP is released
Released eIF2-GDP interacts with
GNF eIF2B & GTP eIF2-GTP for
another round of initiation
Final step: binding of pre-initiation
complex by 60S subunit and
eIF5B-GTP

IRES (internal ribosomal entry sites) = common

in messengers that encode protein that protect
cells from stress, when the translation of most
mRNAs is REPRESSED

Prokaryotes fewer initiation factors

1) 30S-IF3 binds to either mRNA or
ternary complex (IF2 & fMet-tRNA,
GTP)
2) IF1 = also acts in formation of PIC
3) 50S binds to PIC
4) GTP is hydrolyzed release of
initiation factors

Prokaryotes rely more on

RNA-RNA interactions to
position mRNA

ELONGATION

Peptidyl transferase
Growing peptide to alpha-amino group of the
AA residue of the aminoacyl-tRNA
Efficiency & fidelity enhanced by NONribosomal EFs that use GTP
Up to 3 tRNA molecules are bound to specific
sites that span BOTH ribosomal subunits