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Termination
Elongation
Outline
Of all the events we have discussed,translation
is among the most highly conserved across all
organisms and among the most energetically
costly for the cell
Topic 1: mRNA
Polypeptide chains are specified by openreading frames
ORF:the protein coding region of each mRNA is
composed of a contiguous, non-overlapping
string of codons .
Each ORF specifies a single protein and starts
and ends at internal sites within the mRNA .
That is the ends of an ORF are distinct from the
ends of the mRNA.
Two conceptions
Polycistronic mRNA: mRNAs containing multiple
ORFs
usually in prokaryotic cells
Monocistronic mRNA: mRNAs containing a
single ORF
usually in eukaryotic cells
RBS
Prokaryotic mRNAs have a ribosome binding
site that recruits the translational machinery.
RBS:a short sequence upstream of the start
codon in many prokaryotic open-reading frame.
Location:three to nine base pairs on the 5side
of the start codon.
Function:complementary to a sequence locate
near the 3 end of one of the RNA
components ,the 16S ribosomal RNA
One conception:
Translational coupling:
some prokaryotic ORFs internal to a polycistronic message
lack a strong ribosome binding site but are nonetheless
actively translated. In these cases the start codon often
overlaps the 3 end of the adjacent open-reading frame.
Thus ,a ribosome that has just completed translating the
upstream open-reading frame is appropriately positioned to
begin translating from the start codon for the downstream
open-reading frame,circumventing the need for a ribosome
binding site to recruit the ribosome.This phenomenon of linked
translation between overlapping open-reading frames is known
as Translational coupling
Topic 2: tRNA
.tRNAs are adaptors between codons and
amino acids
there are many types of tRNA molecules, but
each is attached to a specific amino acid and
each recognizes a particular codon, or codons.
75~95 ribonucleotides in length.
Features in common:
All tRNAs end at the 3 terminus with the
sequence 5-CCA-3 at the 3 end, where the
aminoacyl tRNA synthetase adds the amino
acid.
The presence of several unusual bases in their
primary structure: U, D and so on
Composed of 3 or
4 bp stem and a
loop called the Dloop (DHU-loop)
usually
containing the
modified base
dihydrouracil.
Consisting of a 5 bp
stem and a 7
residues loop in
which the anticodon
is located. The 3-nt
anticodon sequence
is used to recognize
the codon sequence
in the mRNA
Two steps:
Adenylylation: the amino acid reacts with ATP
to become adenylylated with the concomitant
release of pyrophosphate.
tRNA charging: the adenylylated amino
acid,which remains tightly bound to the
synthetase, reacts with tRNA.
Adenylylation
tRNA charging
How to solve?
tRNA synthetases recognize unique structural
features of cognate tRNAs.
The acceptor stem
The anticodon loop
Ribosome cycles
Polysome/polyribosome
Although a ribosome can synthesize only one
polypeptide at a time, each mRNA can be
translated simultaneously by multiple
ribosomes.
So, an mRNA bearing multiple ribosomes
A question!
Both the decoding center and the peptidyl
transferase center are buried within the intact
ribosome. Yet ,mRNA must be threaded
through the decoding center during translation,
and the nascent polypeptide chain must escape
from the peptidyl transferase center. How do
these polymers enter and exit the robosome?
fMet-tRNAifMet
Key points
EF-G drives translocation by displacing the
tRNA bonnd to the A site.
EF-Tu-GDP and EF-G-GDP must exchange
GDP for GTP prior to participating in a new
round of elongation.
A cycle of peptide bond formation consumes
two molecules of GTP and one molecule of
ATP.
Short regions of
class I release factors
recognize stop codons
and trigger release
of the peptidyl chain.