Transla'on

•  The informa'on in RNA is translated into

proteins, hence, transla'on.

•  mRNA is translated into the amino acid sequence

of a protein by specific rules, known as the
gene'c code.

–  The sequence of nucleo'des in the mRNA molecule is

read in consecu've groups of three (codon)
–  mRNA consists of four different nucleo'des AUGC
–  so there are 43 possible combina'ons of codons
–  But, only 20 amino acids are known !
•  The code is redundant and some amino acids are specified by
more than one codon.

•  Some nucleo'de triplets do not code

Gene'c code is universal

Reading frames

•  An RNA sequence can be

translated in any one of
three different reading
frames.

•  But, only one of the three
possible reading frames in
an mRNA encodes the
required protein.
 Requirements

•  PlaGorm for synthesis •  Ribosomes

•  Template •  mRNA
•  Raw material •  Amino acids
•  carriers •  tRNA (transfer RNA)
 tRNA molecules carry amino acids

•  The codons in an mRNA molecule do not directly

recognize the amino acids

•  The transla'on of mRNA into protein depends on

adaptor molecules known as transfer RNAs
(tRNAs)

•  each about 80 nucleo'des in length.

•  tRNA molecules can fold into precise
3D structures

•  Two regions of unpaired nucleo'des

situated at either end of the molecule
are crucial to the func'on of tRNA in
protein synthesis.

•  One of these regions forms the

an'codon, a set of three consecu've
nucleo'des that pairs with the
complementary codon in an mRNA
molecule.

•  The other is a short single-stranded

region at the 3ʹ end of the molecule;
where the amino acid that matches
the codon is aRached to the tRNA.
Charging of tRNA molecule
•  Each tRNA molecule linked to the one amino acid in 20

•  Recogni'on and aRachment is done by enzymes called

aminoacyl-tRNA synthetases which covalently couple amino
acid to its appropriate tRNA molecule

•  Most cells have a different synthetase enzyme for each amino

acid.

•  synthetase enzymes select the correct amino acid by a two-

step mechanism.

•  The correct amino acid and ATP enters the ac've-site of

synthetase forms AMP-aminoacyl complex

•  The tRNA synthetase recognize the correct set of tRNAs

•  tRNA synthetases directly recognize the matching tRNA

an'codon; these synthetases contain three adjacent
nucleo'de-binding pockets, each of which is complementary
in shape and charge to a nucleo'de in the an'codon.
•  Each ribosome has one
binding site for mRNA in
smaller sub-unit

•  3 binding sites in larger

subunit:
–  A site = aminoacyl-tRNA
–  P site = pep'dyl-tRNA
–  E site = exit
 Ini'a'on of transla'on

•  The site at which protein synthesis begins on the mRNA

is crucial

•  It sets the reading frame for the whole length of the

message.

•  The ini'a'on step is also important because for most

genes it is the last point at which the cell can decide
whether the mRNA is to be translated to produce a
protein.
Ini'a'on of transla'on

•  The transla'on of an mRNA begins with the codon

AUG and an ini'ator tRNA that always carries the
amino acid methionine

•  The ini'ator tRNA is specially recognized by ini'a'on

factors (eIFs) because it has a nucleo'de sequence
dis'nct from that of the tRNA that normally carries
methionine.

•  In eukaryotes, the ini'ator tRNA–methionine complex

(Met–tRNAi) is first loaded into the small ribosomal
subunit along with addi'onal proteins called
eukaryo'c ini'a'on factors, or eIFs.

•  Next, the small ribosomal subunit binds to the 5ʹ end
of an mRNA molecule, which is recognized by virtue of
its 5ʹ cap that has previously bound two ini'a'on
factors, eIF4E and eIF4G

•  The small ribosomal subunit then moves forward along

the mRNA searching for the first AUG
•  At this point, the ini'a'on factors
dissociate allowing the large
ribosomal subunit to assemble with
the complex and complete the
ribosome.

•  The ini'ator tRNA remains at the P

site, leaving the A site vacant.
Protein synthesis is therefore ready
to begin
•  Elonga'on contains 4 major steps:

•  Step 1: A tRNA carrying the next amino acid in
the chain binds to the ribosomal A site by
forming base pairs with the mRNA codon
posi'oned there, so that the P site and the A site
contain adjacent bound tRNAs.

•  Step 2: the carboxyl end of the polypep'de chain
is released from the tRNA at the P site and joined
to the free amino group of the amino acid linked
to the tRNA at the A site, forming a new pep'de
bond. This central reac'on of protein synthesis
is catalyzed by a pep#dyl transferase contained
in the large ribosomal subunit.
•  Step 3:The large subunit moves
rela've to the mRNA held by the
small subunit, thereby shi^ing the
acceptor stems of the two tRNAs to
the E and P sites of the large subunit.

•  Step 4: Conforma'onal changes

moves the small subunit and its
b o u n d m R N A e x a c t l y t h r e e
nucleo'des, ejec'ng the spent tRNA
from the E site and rese_ng the
ribosome so it is ready to receive the
next aminoacyl-tRNA.

•  This four-step cycle is repeated each
'me an amino acid is added to the
polypep'de chain.
 Termina'on
•  The end of the protein-coding message is signalled by
three stop codons (UAA, UAG, or UGA).

•  These are not recognized by a tRNA and do not specify an

amino acid, but instead signal to the ribosome to stop
transla'on.

•  Proteins known as release factors bind to ribosome with a

stop codon posi'oned in the A site, forcing the pep'dyl
transferase in the ribosome to catalyse the addi'on of a
water molecule instead of an amino acid to the pep'dyl-
tRNA.

•  This reac'on frees the carboxyl end of the growing
polypep'de chain from its aRachment to a tRNA molecule,
and since only this aRachment normally holds the growing
polypep'de to the ribosome, the completed protein chain
is immediately released into the cytoplasm.
The structural and func'onal differences between bacterial and eukaryo'c
ribosomes can be exploited for therapy to interfere preferen'ally with the
func'on of bacterial ribosomes.