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Campbell
Shawn O. Farrell
international.cengage.com/
Chapter Twelve
Protein Synthesis: Translation of
the Genetic Message
Chain Initiation
In all organisms, synthesis of polypeptide chain
starts at the N-terminal end, and grows from Nterminus to C-terminus
Initiation requires:
tRNAfmet
initiation codon (AUG) of mRNA
30S ribosomal subunit
50S ribosomal subunit
initiation factors IF-1, IF-2, and IF-3
GTP, Mg2+
Chain Initiation
tRNAmet and tRNAfmet contain the triplet 3-UAC-5
Triplet base pairs with 5-AUG-3 in mRNA
3-UAC-5 triplet on tRNAfmet recognizes the AUG
triplet (the start signal) when it occurs at the beginning
of the mRNA sequence that directs polypeptide
synthesis
3-UAC-5 triplet on tRNAmet recognizes the AUG
triplet when it is found in an internal position in the
mRNA sequence
Start signal is preceded by a Shine-Dalgarno purinerich leader segment, 5-GGAGGU-3, which usually
lies about 10 nucleotides upstream of the AUG start
signal and acts as a ribosomal binding site
Chain Elongation
Uses three binding sites for tRNA present on the
50S subunit of the 70S ribosome: P (peptidyl) site, A
(aminoacyl) site, E (exit) site.
Requires
70S ribosome
codons of mRNA
aminoacyl-tRNAs
elongation factors EF-Tu (Elongation factor
temperature-unstable), EF-Ts (Elongation factor
temperature-stable), and EF-G (Elongation factorGTP)
GTP, and Mg2+
Elongation Steps
Step 1
an aminoacyl-tRNA is bound to the A site
the P site is already occupied
2nd amino acid bound to 70S initiation complex. Defined by the
mRNA
Step 2
EF-Tu is released in a reaction requiring EF-Ts
Step 3
the peptide bond is formed, the P site is uncharged
Step 4
Chain Elongation
Chain Termination
Chain termination requires
stop codons (UAA, UAG, or UGA) of mRNA
RF-1 (Release factor-1) which binds to UAA and
UAG or RF-2 (Release factor-2) which binds to UAA
and UGA
RF-3 which does not bind to any termination codon,
but facilitates the binding of RF-1 and RF-2
GTP which is bound to RF-3
Chain Termination
Protein Synthesis
In prokaryotes, translation begins very soon after
mRNA transcription
It is possible to have several molecules of RNA
polymerase bound to a single DNA gene, each in a
different stage of transcription
It is also possible to have several ribosomes bound to
a single mRNA, each in a different stage of translation
Polysome: mRNA bound to several ribosomes
Coupled translation: the process in which a
prokaryotic gene is being simultaneously transcribed
and translated
Eukaryotic Translation
Chain Initiation:
the most different from process in prokaryotes
13 more initiation factors are given the designation eIF
(eukaryotic initiation factor) (Table 12.4)
Chain termination
stop codons are the same: UAG, UAA, and UGA
only one release factor that binds to all three stop
codons
Posttranslational Modification
Newly synthesized polypeptides are frequently modified
before they reach their final form where they exhibit biological
activity
N-formylmethionine in prokaryotes is cleaved
specific bonds in precursors are cleaved, as for example,
preproinsulin to proinsulin to insulin
leader sequences are removed by specific proteases of the
endoplasmic reticulum; the Golgi apparatus then directs the
finished protein to its final destination
factors such as heme groups may be attached
disulfide bonds may be formed
amino acids may be modified, as for example, conversion of
proline to hydroxyproline
other covalent modifications; e.g., addition of carbohydrates
Protein Degradation
Proteins are in a dynamic state and are often turned
over
Degradative pathways are restricted to
subcellular organelles such as lysosomes
macromolecular structures called proteosomes
Ubiquitin-Proteosome Degradation