GTPase activation of elongation factors Tu and G on the ribosome.
The GTPase activity of elongation factors Tu and G is stimulated by the ribosome. The factor binding site is located on the 50S ribosomal subunit and comprises proteins L7/12, L10, L11, the L11-binding region of 23S rRNA, and the sarcin-ricin loop of 23S rRNA. The role of these ribosomal elements in factor binding, GTPase activation, or functions in tRNA binding and translocation, and their relative contributions, is not known. By comparing ribosomes depleted of L7/12 and reconstituted ribosomes, we show that, for both factors, interactions with L7/12 and with other ribosomal residues contribute about equally and additively to GTPase activation, resulting in an overall 10(7)-fold stimulation. Removal of L7/12 has little effect on factor binding to the ribosome. Effects on other factor-dependent functions, i.e., A-site binding of aminoacyl-tRNA and translocation, are fully explained by the inhibition of GTP hydrolysis. Based on these results, we propose that L7/12 stimulates the GTPase activity of both factors by inducing the catalytically active conformation of the G domain. This effect appears to be augmented by interactions of other structural elements of the large ribosomal subunit with the switch regions of the factors.
D000244:Adenosine Diphosphate / Q000031:analogs & derivatives* / Q000737:chemistry; D001665:Binding Sites; D004789:Enzyme Activation; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000737:chemistry* / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000031:analogs & derivatives* / Q000737:chemistry; D006160:Guanosine Triphosphate / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D009942:Organometallic Compounds / Q000737:chemistry; D020653:Peptide Elongation Factor G / Q000737:chemistry* / Q000378:metabolism; D010444:Peptide Elongation Factor Tu / Q000737:chemistry* / Q000378:metabolism; D010710:Phosphates / Q000737:chemistry / Q000378:metabolism; D021381:Protein Transport; D012360:RNA, Transfer, Phe / Q000737:chemistry; D012269:Ribosomal Proteins / Q000737:chemistry / Q000378:metabolism; D012270:Ribosomes / Q000201:enzymology*; D062367:ortho-Aminobenzoates / Q000737:chemistry
D029968:Escherichia coli Proteins; D009942:Organometallic Compounds; D020653:Peptide Elongation Factor G; D010710:Phosphates; D012360:RNA, Transfer, Phe; D012269:Ribosomal Proteins; C105164:adenosine diphosphate tetrafluoroaluminate; D062367:ortho-Aminobenzoates; C065197:3'-(methylanthraniloyl)-2'-deoxy-guanosine diphosphate; D006153:Guanosine Diphosphate; D000244:Adenosine Diphosphate; C016700:ribosomal protein L7-L12; D006160:Guanosine Triphosphate; D020558:GTP Phosphohydrolases; D010444:Peptide Elongation Factor Tu