Clustering and co-transcription of the Bacillus subtilis genes encoding the aminoacyl-tRNA synthetases specific for glutamate and for cysteine and the first enzyme for cysteine biosynthesis.
The Bacillus subtilis cysE and cysS genes encoding, respectively, the serine acetyltransferase and the cysteinyl-tRNA synthetase were found downstream from the gltX gene encoding the glutamyl-tRNA synthetase. This gene organization is also conserved in Bacillus stearothermophilus where the cysE and cysS genes show high amino acid identity with those of B. subtilis. In both organisms the coding sequences of cysE and cysS overlap, suggesting a translational coupling. B. subtilis cysE and cysS were expressed in Escherichia coli using the inducible trc promoter; they functionally complement mutants of E. coli affected in those genes. Overproduction of B. subtilis CysRS in E. coli has a toxic effect on cell growth. Disruption of gltX and cysS by Campbell-type insertion is lethal for the cell, indicating that these genes code for an essential and unique function in B. subtilis. S1 mapping analysis shows that the transcription of gltX is under the control of a sigma A promoter located 43 base pairs upstream of the initiation codon. A T-box sequence and a rho-independent terminator known to regulate expression of other aminoacyl-tRNA synthetase genes and of some amino acid biosynthetic operons in Bacillus sp., were found between gltX and cysE. No sigma A promoter was detected upstream of cysE, which is consistent with the lethality of a Campbell-type insertion using a plasmid that interrupts transcription coming from the gltX promoter, and suggests that gltX, cysE, and cysS constitute an operon. This is the first case where genes implicated in the biosynthesis of an amino acid and its cognate aminoacyl-tRNA synthetase are shown to be co-transcribed.
Gagnon
Y
Y
Département de Biochimie, Faculté des Sciences et de Génie, Université Laval, Québec, Canada.
Breton
R
R
Putzer
H
H
Pelchat
M
M
Grunberg-Manago
M
M
Lapointe
J
J
D000595:Amino Acid Sequence; D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics* / Q000378:metabolism; D001412:Bacillus subtilis / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D003545:Cysteine / Q000096:biosynthesis / Q000378:metabolism*; D004269:DNA, Bacterial; D004926:Escherichia coli / Q000235:genetics; D001411:Geobacillus stearothermophilus / Q000235:genetics; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D008969:Molecular Sequence Data; D005810:Multigene Family; D009690:Nucleic Acid Conformation; D012329:RNA, Bacterial / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014158:Transcription, Genetic
D004269:DNA, Bacterial; D005971:Glutamates; D012329:RNA, Bacterial; D018698:Glutamic Acid; D000604:Amino Acyl-tRNA Synthetases; C019975:cysteinyl-tRNA synthetase; D003545:Cysteine