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BIO353 Lecture11 (Translation) SHF 2021 Web
BIO353 Lecture11 (Translation) SHF 2021 Web
Langenscheidt
Standard Dictionary
Protein
DNA - RNA
RNA - AA
shf – 2021
Translation: general overview - central dogma
shf – 2021
Translation: basic ingredients
processed mRNA
ribosome
protein synthesis
shf – 2021
Translation: general overview
shf – 2021
Translation: general overview
shf – 2021
Translation: general overview
AUG AAA GCA AUU UUC GUA CUG AAA GGU UGG UGG UGA
Frame 1
M K A I F V L K G W W stop
A UGA AAG CAA UUU UCG UAC UGA AAG GUU GGU GGU GA
Frame 2
stop K Q F S Y stop K V G G
AU GAA AGC AAU UUU CGU ACU GAA AGG UUG GUG GUG A
Frame 3
E S N F R T E R L V V
shf – 2021
Translation: initiation - prokaryotes vs eukaryotes
polycistronic gene
cistron 1 cistron 2 cistron 3
(= ORF 1) (= ORF 2) (= ORF 3)
shf – 2021
Translation: initiation in prokaryotes – Shine-Dalgarno sequence
araB UUUGGAUGGAGUGAAACGAUGGCGAUU
gelE AGCCUAAUGGAGCGAAUUAUGAGAGUU
lacI CAAUUCAGGGUGGUGAUUGUGAAACCA
lacZ UUCACACAGGAAACAGCUAUGACCAUG
trpE CAAAAUUAGAGAAUAACAAUGCAAACA
trpL GUAAAAAGGGUAUCGACAAUGAAAGCA
talA UUUCAAGGAGUAUUUCCUAUGAACGAG
dsdX CACAGGGGAAGGUGAGAUAUGCACUCU
shf – 2021
Translation: initiation in prokaryotes – Shine-Dalgarno sequence
araB UUUGGAUGGAGUGAAACGAUGGCGAUU
gelE AGCCUAAUGGAGCGAAUUAUGAGAGUU
lacI CAAUUCAGGGUGGUGAUUGUGAAACCA
lacZ UUCACACAGGAAACAGCUAUGACCAUG
trpE CAAAAUUAGAGAAUAACAAUGCAAACA
trpL GUAAAAAGGGUAUCGACAAUGAAAGCA
talA UUUCAAGGAGUAUUUCCUAUGAACGAG
dsdX CACAGGGGAAGGUGAGAUAUGCACUCU
shf – 2021
Translation: initiation in prokaryotes – Shine-Dalgarno sequence
ribosomal RNA
Shine-Dalgarno sequence
polycistronic gene
shf – 2021
Translation: initiation - prokaryotes vs eukaryotes
ribosomal RNA
Shine-Dalgarno sequence
polycistronic gene
Kozak sequence
monocistronic gene
shf – 2021
Translation: tRNA structure and function
amino acid
shf – 2021
Translation: tRNA structure and function
à no degradation (dihydrouridine)
shf – 2021
Translation: and the genetic code(s)
shf – 2021
Translation: charging (= loading) tRNAs with amino acids
shf – 2021
Translation: charging tRNAs with amino acids - step 1
adenylylation
shf – 2021
Translation: charging tRNAs with amino acids - step 2
tRNA charging
charged tRNA
shf – 2021
Translation: charging tRNAs with amino acids - tRNA synthetases
AA à 3’-OH AA à 2’-OH
shf – 2021
Translation: charging tRNAs with amino acids - tRNA synthetases
Aminoacyl-tRNA
synthetase
amino acid
ATP
tRNA
AMP + 2 Pi
charged
tRNA
shf – 2021
Translation: charging tRNAs with amino acids - tRNA synthetases
discriminator base
Aminoacyl-tRNA tRNA
synthetase
amino acid
ATP
amino acid
tRNA
AMP + 2 Pi
charged anticodon
tRNA
tRNA synthetase
shf – 2021
Translation: charging tRNAs with amino acids - tRNA synthetases
shf – 2021
Translation: charging tRNAs with amino acids - tRNA synthetases
shf – 2021
Translation: the structure of the ribosome
Theodor Svedberg
(1884 – 1971)
shf – 2021
Translation: the structure of the ribosome
Kaltschmidt, E. and H.G. Wittmann, Ribosomal proteins XII: Number of proteins in small and large ribosomal subunits of Escherichia coli as
determined by two-dimensional gel electrophoresis. PNAS USA, (1970, 67:1277–78.)
shf – 2021
Translation: the structure of the ribosome
prokaryotes
16S rRNA
shf – 2021
Translation: the structure of the ribosome
prokaryotes
peptidyl-transferase center
decoding center
shf – 2021
Translation: the structure of the ribosome
eukaryotes prokaryotes
shf – 2021
Translation: the ribosome cycle - prokaryotes
shf – 2021
Translation: polyribosomes (polysomes)
prokaryotes cotranscriptional
in cytoplasm
eukaryotes
30 nt
~80 nt
shf – 2021
Translation: polypeptide synthesis reaction
amino acid
amino acid
peptidyl transferase
reaction
shf – 2021
Translation: the ribosome structure - EPA sites
shf – 2021
Translation: the ribosome structure - EPA sites
decoding center
peptidyl transferase
mostly rRNA
center
devoid of protein
shf – 2021
Translation: the ribosome structure - EPA sites
P A
P
E A
AA AA
shf – 2021
Translation: the ribosome structure - EPA sites
large subunit
A
P
E
shf – 2021
Translation: initiation in prokaryotes
identification of
initiation translation start codon
shf – 2021
Translation: initiation in prokaryotes
step 1 step 2
binding of binding of
mRNA and initiator tRNA large ribosomal subunit
(any order)
shf – 2021
Translation: initiation in prokaryotes - start site
shf – 2021
Translation: initiation in prokaryotes - start site
initiator tRNA
vs
shf – 2021
Translation: initiation in prokaryotes - start site
shf – 2021
Translation: initiation in prokaryotes - regulatory factors
dissociation of
the small subunit
occupation of
E site
shf – 2021
Translation: initiation in prokaryotes - regulatory factors
occupation of
A site
dissociation of
the small subunit initiator tRNA
loading to P site
occupation of
E site
shf – 2021
Translation: initiation in prokaryotes - regulatory factors
binding of
large subunit
factor binding
release of IF3 center
release of IF1 / 2
(GTP hydrolysis by IF2)
shf – 2021
Translation: initiation in eukaryotes - regulatory factors
ribosome
dissociation of
the small subunit
occupation of
E and A site
Met-tRNAMet
loading to P site
43S preinitiation
complex
shf – 2021
Translation: initiation in eukaryotes - regulatory factors and events
ribosome mRNA
recognition of 5’-CAP
helicase activity /
removal of secondary structure
43S preinitiation
complex
shf – 2021
Translation: initiation in eukaryotes - regulatory factors and events
ribosome mRNA
re-initiation
shf – 2021
Translation: initiation in eukaryotes - scanning
scanning à
shf – 2021
Translation: initiation in eukaryotes - subunit recruitment
shf – 2021
Translation: initiation in eukaryotes - subunit recruitment
eviction of IFs
à elongation
shf – 2021
Translation: initiation in eukaryotes - internal ribosome entry sites (IRES)
shf – 2021
Translation: elongation - steps
shf – 2021
Translation: elongation - steps
shf – 2021
Translation: elongation - steps
shf – 2021
Translation: elongation - proofreading (EF-Tu)
1. - proofreading by 2. - proofreading by
16S rRNA EF-Tu
shf – 2021
Translation: elongation - proofreading (accommodation)
3. - proofreading by
accommodation
shf – 2021
Translation: elongation - translocation
shf – 2021
Translation: elongation - translocation
shf – 2021
Translation: elongation - translocation
shf – 2021
Translation: termination - release factors
t-RNA
release factor
shf – 2021
Translation: termination - release factors
shf – 2021
Translation: termination - release factors
shf – 2021
Translation: termination - ribosome recycling
shf – 2021
Translation: what if something goes wrong? - no STOP codon
shf – 2021
Translation: what if something goes wrong? - no STOP codon
shf – 2021
Translation: what if something goes wrong? - no STOP codon
shf – 2021
Translation: what if something goes wrong? - stalling
shf – 2021
Translation: what if something goes wrong? - no STOP codon
shf – 2021
Translation: what if something goes wrong? - premature STOP codon
nonsense-mediated decay
shf – 2021
Translation: what if something goes wrong? - premature STOP codon
nonsense-mediated decay
shf – 2021