Characterization of altered forms of glycyl transfer ribonucleic acid synthetase and the effects of such alterations on aminoacyl transfer ribonucleic acid synthesis in vivo.
The glycyl transfer ribonucleic acid (tRNA) synthetase (GRS) activities of several Escherichia coli glyS mutants have been partially characterized; the K(m) for glycine and the apparent V(max) of several of the altered GRS differ significantly from the parental GRS. Paradoxically, some of the altered forms exhibit more activity in vitro than the GRS from a prototrophic strain (GRS(L)); several parameters of these activities have been studied in an attempt to resolve this problem. The amount of acylated tRNA(Gly) in vivo was examined to assess the GRS activities inside the cells. During exponential growth in media containing glycine, moderate amounts of acylated tRNA(Gly) occur in the glyS mutants; glycine deprivation leads to a dramatic drop in the amount of acylated tRNA(Gly). An alternative measure of the in vivo activities of the altered enzymes is the efficiency of suppression of the trpA36 locus by su(36) (+); glyS mutants grown with added glycine exhibit one-third to one-fourth the suppression efficiency of the prototrophic glyS(H) parent, presumably because they are less efficient, even in the presence of high levels of glycine, in charging the tRNA(Gly) species which functions as the translational suppressor.
D000596:Amino Acids; D002247:Carbon Isotopes; D002498:Centrifugation; D002848:Chromatography, DEAE-Cellulose; D003470:Culture Media; D004926:Escherichia coli / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D005827:Genetics, Microbial; D005998:Glycine; D007211:Indoles; D008025:Ligases / Q000378:metabolism*; D009154:Mutation; D012343:RNA, Transfer / Q000096:biosynthesis*; D013696:Temperature; D014316:Tritium; D014364:Tryptophan
D000596:Amino Acids; D002247:Carbon Isotopes; D003470:Culture Media; D007211:Indoles; D014316:Tritium; D014364:Tryptophan; D012343:RNA, Transfer; D008025:Ligases; D005998:Glycine