Protein Synthesis

Protein Synthesis

OVERVIEW
Genetic information in chromosomes
transmitted to daughter cells (DNA
replication)
- expressed to RNA (transcription)
- protein synthesis (translation).

Protein Synthesis

Protein Synthesis
THE GENETIC CODE
Dictionary identify the
correspondence between a
seq of nucleotide bases & seq
of a.a.
Each individual word in the
code composed of 3
nucleotide bases = codon

Protein Synthesis
Codons
-presented in mRNA language of A, G, C, U
-written from 5 to 3-end
-used to produced three-base codons
-64 combinations of bases
-code for 20 common a.a.

Protein Synthesis
How to translate a codon?
Table
Termination (stop or nonsense) codons
UAG, UGA, UAA

Protein Synthesis

Protein Synthesis
Characteristics of the genetic code
1. Specificity
-specific, particular codon always codes the same a.a.
2. Universality
-conserved
3. Degeneracy
-degenerate/redundant (a given a.a. may have more than
1 triplet coding for it. i.e Arg by 6 different codons
4. Nonoverlapping & commaless
-code is read from fixed starting point as continuous seq
of bases, taken 3 at a time

Protein Synthesis
Consequences of altering the nucleotide sequence
Silent mutation
Missense mutation

Nonsense mutation
Other mutations
Trinucleotide repeat expansion
Splice site mutations
Frame-shift mutations

Protein Synthesis

Protein Synthesis

Protein Synthesis

Protein Synthesis
COMPONENTS REQUIRED FOR TRANSLATION
1. Amino acids
2. Transfer RNA
3. Aminoacyl-tRNA
4. Messenger RNA
5. Functionally competent ribosomes
6. Protein factors
7. ATP & GTP

Protein Synthesis
COMPONENTS REQUIRED FOR TRANSLATION
1. Amino acids

Protein Synthesis
COMPONENTS REQUIRED FOR TRANSLATION
2. tRNA (carry specific a.a. & to recognize the codon for that
a.a. / adaptor molecules)
-1 specific type of tRNA is req per a.a.
-Amino acid attachment site
attachment site for specific a.a at 3
-Anticodon
3-base nucleotide seq. that
recognizes specific codon on
mRNA

Protein Synthesis

Protein Synthesis
COMPONENTS REQUIRED FOR TRANSLATION
3. Aminoacyl-tRNA synthetases
enz req for attachment of a.a. to their corresponding
tRNA

Protein Synthesis
COMPONENTS REQUIRED FOR
TRANSLATION
Catalyzes the esterification of a
specific a. a. or its precursor to one of
all its compatible cognate tRNAs to
form an aminoacyl-tRNA

This sometimes called "charging"

tRNA with a. a.
Once the tRNA is charged, a ribosome
can transfer the a. a. from the tRNA
onto a growing peptide, according to
the genetic code.

Protein Synthesis
COMPONENTS REQUIRED FOR TRANSLATION
4. mRNA
specific mRNA req as template for polypeptide
synthesis

Protein Synthesis
COMPONENTS REQUIRED FOR TRANSLATION
5. Functionally competent ribosomes
ribosomes = protein + ribosomal RNA complexes
2 subunits = large & small
large catalyzes formation of peptide bonds
small binds mRNA

Protein Synthesis
COMPONENTS REQUIRED FOR
TRANSLATION
3 sites:
A site = point of entry for the
aminoacyl tRNA
P site = where the peptidyl tRNA is
formed in the ribosome
E site = the exit site of uncharged
tRNA after it gives its
amino acid to the
growing peptide chain

Protein Synthesis
COMPONENTS REQUIRED FOR TRANSLATION
6. Protein factors
Initiation, Elongation & Termination factors
7. ATP & GTP
sources of energy

Protein Synthesis
STEPS IN PROTEIN SYNTHESIS
Translation
Initiation
Elongation
Termination
Polypeptide chains produced may be modified by
posttranslational modification

Protein Synthesis
1. Initiation
2 ribosomal subunits
mRNA,
aminoacyl-tRNA,
GTP / ATP,
Initiation factors
Prokaryotes IFs : IF-1, IF-2, IF-3

Eukaryotes IFs : >10, eIF

Protein Synthesis
1. Initiation
2 mechanisms ribosome recognizes nucleotide:
i) Shine-Dalgarno seq. : E. coli, 6-10 bases upstream
AUG
Euk: 40S ribosomal subunit binds to cap structure at
5 end of mRNA & moves until AUG
ii) Initiation codon AUG recognized by initiator tRNA
Euk : not formylated

Protein Synthesis
1. Initiation
1st aminoacyl tRNA (fMet-tRNA)

Protein Synthesis
1. Initiation

Protein Synthesis

Initiation
Inhibitors of translation

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2. Elongation
Elongation Factors
EF-Tu, EF-Ts
EF-1 , EF-1

Translocation
EF-G
EF-2

Protein Synthesis

2. Elongation
Addition of a.a. to carboxyl end of growing chain

Ribosome moves from 5-end to 3end of mRNA

Protein Synthesis

Protein Synthesis
3. Termination
When 1 of 3 termination codons moves into A site
Release Factors
RF-1 : UAA, UAG ; RF-2 : UGA, UAA ; RF-3 : release
RF-1 & RF-2
eRF : recognizes all termination codons ; eRF-3

Protein Synthesis
3. Termination

Protein Synthesis
Polysomes
> 1 ribosome at a time can generally translate a message
Complex of 1 mRNA + a number of ribosomes = polysome /
polyribosome

Protein Synthesis

Protein Synthesis

Protein Synthesis