The Escherichia coli GTPase CgtAE cofractionates with the 50S ribosomal subunit and interacts with SpoT, a ppGpp synthetase/hydrolase.
CgtA(E)/Obg(E)/YhbZ is an Escherichia coli guanine nucleotide binding protein of the Obg/GTP1 subfamily whose members have been implicated in a number of cellular functions including GTP-GDP sensing, sporulation initiation, and translation. Here we describe a kinetic analysis of CgtA(E) with guanine nucleotides and show that its properties are similar to those of the Caulobacter crescentus homolog CgtA(C). CgtA(E) binds both GTP and GDP with moderate affinity, shows high guanine nucleotide exchange rate constants for both nucleotides, and has a relatively low GTP hydrolysis rate. We show that CgtA(E) is associated predominantly with the 50S ribosomal subunit. Interestingly, CgtA(E) copurifies with SpoT, a ribosome-associated ppGpp hydrolase/synthetase involved in the stress response. The interaction between CgtA(E) and SpoT was confirmed by reciprocal coprecipitation experiments and by two-hybrid assays. These studies raise the possibility that the ribosome-associated CgtA(E) is involved in the SpoT-mediated stress response.
Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, 48109, USA.
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Wout
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Sullivan
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Reese
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Zhou
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Lin
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Maddock
D001426:Bacterial Proteins; D016935:Caulobacter crescentus / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry* / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000302:isolation & purification / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D008025:Ligases / Q000302:isolation & purification / Q000378:metabolism*; D020559:Monomeric GTP-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding; D025941:Protein Interaction Mapping; D012270:Ribosomes / Q000737:chemistry*; D020798:Two-Hybrid System Techniques
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D006153:Guanosine Diphosphate; D006160:Guanosine Triphosphate; C108621:CgtA protein, bacteria; C491572:ObgE protein, E coli; D020559:Monomeric GTP-Binding Proteins; D008025:Ligases; C028812:guanosine 3',5'-polyphosphate synthetases