Translation

Ribosomes synthesize a polypeptide according

to the genetic instructions in mRNA.
Translation in Eukaryotes occurs in 3 steps
1. Initiation
• Ribosome binds to the mRNA and initiates at an AUG
(methionine) codon

2. Elongation
• The polypeptide is lengthened one amino acid at a time

3. Termination
• Synthesis of the polypeptide terminates and the ribosome
dissociates from the mRNA and the polypeptide
 Initiation of Translation (Eukaryotes)
• The small ribosomal
subunit forms an
initiation complex with
the initiator tRNA.

• This complex binds the 5’

cap, then scans 5’ to 3’ in
search of the first AUG
codon.
Initiation of Translation
(Eukaryotes)
• Once the first AUG is
located then the large
ribosomal subunit to binds to
form the functional ribosome
• An aminoacyl-tRNA then
binds to the A site and a
peptide bond is formed
between methionine and the
amino acid in the A site
• Initiation is now complete
and elongation can proceed
 Elongation
• The nascent (growing) polypeptide
is bound to the peptidyl-tRNA in
the P-site

• The incoming aminoacyl-tRNA

binds to the A-site

• Peptidyl transferase catalyzes a

peptide bond between the new
amino acid and the nascent
polypeptide and breaks the bond
between the nascent chain and the
peptidyl-tRNA

• The ribosome translocates by one

codon along the mRNA
 Elongation
• The tRNA in the E site
exits the ribosome

• The A site receives the next

incoming aminoacyl-tRNA

• A peptide bound is formed

and the cycle continues
until termination
 Termination
• Releasing factors bind to
termination codons (UAG,
UAA, UGA) in the A-site.

• Releasing factors facilitate

hydrolysis of the nascent
polypeptide from the
peptidyl-tRNA thus
freeing the polypeptide
from the ribosome.
 Termination
• Release of the polypeptide
is followed by dissociation
of the ribosomal subunits
from the mRNA
 The 5’ cap and poly-A tail increase translation rates.
• Specialized proteins bind to the
5’cap and 3’ poly-A tail of the
mRNA. These proteins then
bind to each other and thus
bring the 5’ and 3’ ends of the
mRNA together, forming a
circle. Ribosomes that terminate
translation are physically close
to the 5’ cap where they bind
and begin translation again. In
this way the 5’ cap and poly-A
tail function to increase
translation rates.
• Many ribosomes translate the
same mRNA simultaneously.
This complex is a polyribosome
(polysome)
 Post Translational Modifications
• The process of gene
expression is not finished
when an mRNA has been
translated.
• Many post translational
modifications may be required
for a polypeptide to fold into
the shape required for
function.
 Post Translational Modifications
• Modifications may include:
– Phosphorylation, the addition of a
phospahate group
– Methylation, the addition of a
methyl group
– Glycosylation, the addition of
sugar groups
– Disulfide bonds, the formation of
covalent bonds between 2 cysteine
amino acids.
– Proteolytic Cleavage, the cutting
of a sequence of amino acids from
the polypeptide
– Subunit binding to form a
multisubunit protein
Protein Folding begins during Translation
• A polypeptide begins
to fold as soon as it
leaves the ribosome.
• Some polypeptides
can fold into their
complete, mature
conformation without
help.
• However, most
polypeptides required
chaperones to help
them fold properly.
 Heat Shock Proteins (HSP) acts as
Chaperones that aid protein folding
• Chaperones bind to hydrophobic regions of the
polypeptide and shield them from the aqueous environment
until the entire polypeptide is translated. Then the
chaperones help the protein to fold into its proper shape,
with the hydrophobic R groups in the interior of the protein
 Misfolded proteins are destroyed by
Proteosomes
• The proteosome is a barrel-
shaped, multisubunit
protease. Misfolded proteins
enter one end and come out
the other as small chains of
amino acids (peptides) that
are ultimately recycled.
• Proteosomes are abundant,
making up 1% of total
cellular protein.
Ubiquitin binds to misfolded
proteins, targeting them for
destruction by the proteosome

•Misfolded proteins and proteins with

oxidized or abnormal amino acids are
seen as abnormal by the cell.
Specialized enzymes attach chains of
ubiquitin to these abnormal proteins.
Ubiquitin has affinity for the
proteasome and thus brings the
abnormal protein to the proteasome
for destruction. Almost without
exception, proteins that enter the
proteosome are first bound to
ubiquitin.
 Plasma membrane proteins and secreted proteins
are post-translationally modified in the Rough
Endoplasmic Reticulum (RER) and the Golgi
•Proteins bound for the plasma
membrane must first enter the
RER where they begin to fold and
undergo chemical modification.
•Proteins leave the RER in
vesicles, which then fuse with the
golgi, delivering their protein
cargo.
•Proteins are further modified in
the golgi before being sent in
vesicles to the plasma membrane
where they are either secreted or
integrated into the membrane.
 A signal sequence directs a nascent polypeptide to
the rough endoplasmic reticulum
• Only proteins that have an
N-terminal signal sequence
can enter the ER.
• A signal recognition
protein(SRP) binds to the
signal sequence as it first
exits the ribosome and
brings the ribosome to the
ER.
• The polypeptide is threaded
through the ER membrane
and into the ER lumen
A signal sequence directs a nascent polypeptide
to the rough endoplasmic reticulum
A signal sequence directs a nascent polypeptide
to the rough endoplasmic reticulum
 A signal sequence directs a nascent polypeptide to
the rough endoplasmic reticulum
• Once the polypeptide is
completely threaded into
the ER, the signal
sequence is cleaved off .
• Now the polypeptide
undergoes post-
translational modification
and begins to fold
 Protein glycosylation occurs in the ER and
the Golgi
• Most proteins that enter the ER
will be glycosylated, which
means that an oligosaccharide
(branched sugar group) will be
covalently attached to the
protein.
• The sugar group may ultimately
be important to the proteins
function or it may simply act as
an address label required to get
the protein to its next cellular
destination.