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Initiation
Elongation
Termination
Stages of translation
Initiation
_______________________________
prokaryotes: mRNA Shine-Delgarno sequence which hydrogen bonds
with rRNA
eukaryotes: ribosome attaches to 5 cap
Translation Initiation
Initiation requirements:
mRNA molecule
______________________
Initiator tRNA
Three initiation factors
GTP
Mg2+
Initiation
Small ribosomal subunit binds to
_______________________________________
This complex binds to mRNA
In prokaryotes initiation codon _____________ calls for
f-met (N-formylmethionine)
Shine-Dalgarno sequence AGGAGG precedes start
codon in mRNA; complementary sequence on 3of 16S
rRNA
________________________________________
Initiation protein enhances binding of f-met tRNA to the
small subunit sets the reading frame initiation
complex
TRANSLATION INITIATION:
Bacteria
5
Ribosome
binding site
AGGAGGU
Initiation
codon
AUG
3-10 nucleotides
Translation factors:
-initiation factors (IF-1, IF-2, IF-3)
-elongation factors (EF-Tu, EF-Ts,EF-G)
-release factors (RF-1, RF-2, RF-3)
-ribosome recycling factor (RRF)
Initiation Complex
30S small subunit binds
_____________________
30S +IFs binds to mRNA AUG codon region
A sequence upstream of AUG is necessary, -8-10
position
5end of 5UTR ribosome-binding site RBS
(Shine-Delgarno sequence AGGAG purinerich, complementary to UCCUCC pyrimidine
rich region at 3-end of 16S rRNA)
Initiation Complex
Initiator tRNA binds to AUG start codon
Initiator fMet
Methionyl-tRNA synthetase add methionine to
tRNA
Transformylase adds formyl group to methionine
fMet-tRNA.fMet
Initiation Complex
fMet-tRNA binds to AUG of 30S-mRNA complex,
IF3 is released
50S subunit binds
GTP hydrolysis
IF1, IF2 released
Elongation In Prokaryotes
Anticodon of fMet-tRNA is hydrogen-bonded to
AUG in P site
Next codon is in A site
Complementary tRNA binds to A site
tRNA is bound to protein elongator factor EF-Tu
aa-tRNA binds to A site, GTP hydrolysis,
________________________________________
Elongation
Aminoacyl tRNA binds to A site
Growing chain of polypeptide is transferred from
tRNA in P site to tRNA in A site by forming a new
peptide bond
Ribosome translocate along mRNA to position
next codon against A site
tRNA translocate from A-P to P-E sites
Elongation factor EF-TuGTP provide energy
Translocation
Termination
Chain-termination codons UAA, UAG, UGA enter A
site
Recognized by release factors RFs (Rf1 and Rf2; only
one in eukaryotes eRF)
Presence of release factor causes the activity change
of peptidyl transferase, add water to carboxyl of
polypeptide chain
Reaction releases polypeptide from tRNA in the P site
Free tRNA moves to E site, mRNA released, ribosome
dissociate
Termination
,
Ribosome Recycling Factor
Elongation Factor G (EF-G)
function to release mRNA and tRNAs from
ribosomes and dissociate the 70s ribosomes
into the 30s and 50s subunits.
Termination
IF-3 also helps the ribosome-recycling
process by converting transiently
dissociated subunits into stable ones by
binding to ________________________.
This "recycles" the ribosomes for additional
rounds of translation
Ribozymes
A catalytic RNA RNA enzyme
16S rRNA catalysis of peptide bond
Release factor RNA molecule similar to
tRNA
Ribonuclease P - that cleaves the head (5')
end of the precursors of transfer RNA
(tRNA) molecules
snRNPs?
Translation In Eukaryotes
Similar, more ______________________
tRNA Met is not fMet tRNAs coded by
two different genes
Initiation complex starts at AUG closest to
the 5end
Optimal initiation sequence 5GCC(A/G)CCAUGC-3
G increase efficiency, Kozak rules
Translation In Eukaryotes
tRNAiMet + IF----- P site directly
5cap binding protein CBP (part of eIF-4F initiation
complex) binds to 7-metG at 5end on mRNA
Other IFs bind to CBP-mRNA complex joined by
40S subunit of ribosome
Initiation complex moves 5to 3searching for AUG
( almost always first AUG at 5end)
AUG found IFs released ( except eIF-4F), 60S
binds, complete 80S ribosome
Translation In Eukaryotes
80S initiation complex with Met-tRNA
bound to mRNA with AUG in P site
polyA tail is bound by poly(A)binding
protein (PABP), binds to eIF-4G
Initiation of translation is stimulated
Translation
Prokaryotes
ribosomes are smaller
translation is coupled to
transcription
more than one gene may be
translated from the same
mRNA
translation is polycistronic
the Shine-Delgarno sequence
aligns mRNA on ribosome
the first amino acid
incorporated is Nformylmethionine
Eukaryotes
translation monocistronic
ribosomes recognize
mRNA by binding to 7methylguanosine cap at 5'
end of mRNA
methionine is always the
first amino acid
incorporated
Mutations
Mutations
Change in genetic material -a mutation is an
________________________________.
Source of genetic variability
Novel phenotype ___________________
Mutations
Change in chromosome ________________
Change in chromosome ________________
Change in the structure of individual gene
_________________________
Mutations
Somatic or germinal
Spontaneous or induced
Reversible
Gain of function, loss of function, neutral
Point Mutations
Addition
Deletion
Substitution
Frame-shift
Missense
Nonsense
Neutral
silent