Nonribosomal synthesis of guanosine 5',3'-polyphosphates by the ribosomal wash of stringent Escherichia coli.
A factor in the ribosomal wash of stringent strains of E. coli was identified by Haseltine et al. as a complement to the ribosomal system for the synthesis of the magic spot compounds of Cashel and Gallant. This factor has been found, in the absence of ribosomes, to catalyze the enzymatic pyrophosphoryl transfer from ATP to GTP or GTP in the formation of magic spot I and magic spot II, the guanosine tetra- and pentaphosphates (ppGpp and pppGpp), respectively. The enzyme, which normally requires the presence of the ribosome-tRNA-mRNA complex for activity, catalyzes a very slow synthesis that is stimulated tenfold by 20% methanol. The temperature optimum of the methanol-stimulated system is 25-30 degrees and activity is drastically depressed at 37 degrees , presumably by inactivation. Catalysis is linear with enzyme concentration and with time for the first 3 hr; during this period 25% of the added GTP is converted. The nonribosomal system is distinguished from the ribosomal system by having a lower Mg(++) and a higher NH(4) (+) optimum. The two systems differ in their response to antibiotics: thiostrepton strongly inhibits the ribosomal system but has no effect on the nonribosomal system.
D000643:Ammonium Chloride; D004926:Escherichia coli / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D006150:Guanine Nucleotides / Q000096:biosynthesis*; D006160:Guanosine Triphosphate / Q000378:metabolism; D007700:Kinetics; D008274:Magnesium; D000432:Methanol; D010759:Phosphorus Isotopes; D010770:Phosphotransferases / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism*; D013696:Temperature
D006150:Guanine Nucleotides; D010759:Phosphorus Isotopes; D000643:Ammonium Chloride; D006160:Guanosine Triphosphate; D010770:Phosphotransferases; D008274:Magnesium; D000432:Methanol