Elongation factor Tu, a GTPase triggered by codon recognition on the ribosome: mechanism and GTP consumption.
The mechanism of elongation factor Tu (EF-Tu) catalyzed aminoacyl-tRNA (aa-tRNA) binding to the A site of the ribosome was studied. Two types of complexes of EF-Tu with GTP and aa-tRNA, EF-Tu.GTP-aa-tRNA (ternary) and (EF-Tu.GTP)2.aa-tRNA (quinternary), can be formed in vitro depending on the conditions. On interaction with the ribosomal A site, generally only one molecule of GTP is hydrolysed per aa-tRNA bound and peptide bond formed. The second GTP molecule from the quinternary complex is hydrolyzed only during translation of an oligo(U) tract in the presence of EF-G. The first step in the interaction between the ribosome and the ternary complex is the codon-independent formation of an initial complex. In the absence of codon recognition, the aa-tRNA-EF-Tu complex does not enter further steps of A site binding and remains in the initial binding state. Despite the rapid formation of the initial complex, the rate constant of GTP hydrolysis in the noncognate complex is four orders of magnitude lower compared with the cognate complex. This, together with the results of time-resolved fluorescence measurements, suggests that codon recognition by the ternary complex on the ribosome initiates a series of structural rearrangements that result in a conformational change of EF-Tu, presumably involving the effector region, which, in turn, triggers GTP hydrolysis and the subsequent steps of A site binding.
Rodnina
M V
MV
Institute of Molecular Biology, University Witten/Herdecke, Germany.
Pape
T
T
Fricke
R
R
Wintermeyer
W
W
D001483:Base Sequence; D002384:Catalysis; D003062:Codon; D020963:GTP Phosphohydrolase-Linked Elongation Factors / Q000235:genetics*; D006160:Guanosine Triphosphate / Q000378:metabolism*; D008969:Molecular Sequence Data; D010444:Peptide Elongation Factor Tu / Q000235:genetics*; D012345:RNA, Transfer, Amino Acid-Specific / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D003062:Codon; D012345:RNA, Transfer, Amino Acid-Specific; D006160:Guanosine Triphosphate; D020963:GTP Phosphohydrolase-Linked Elongation Factors; D010444:Peptide Elongation Factor Tu