Open AccessCharacterization of the relA/spoT gene from Bacillus stearothermophilus 1
Author(s) -
Wendrich Thomas M.,
Beckering Carsten L.,
Marahiel Mohamed A.
Publication year - 2000
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2000.tb09286.x
Subject(s) - bacillus subtilis , biology , gene , nucleic acid sequence , microbiology and biotechnology , heterologous expression , stringent response , genetics , peptide sequence , bacillaceae , biochemistry , recombinant dna , escherichia coli , bacteria
By use of degenerate primers, we amplified a fragment of a relA/spoT homologous gene from Bacillus stearothermophilus . Chromosomal walking enabled us to sequence the entire gene and its flanking regions. The primary sequence of the gene product is 78% identical to the RelA/SpoT homologue of Bacillus subtilis , and both gene loci share a similar genetic organization. The B. stearothermophilus rel gene was analyzed in vivo by heterologous expression in the B. subtilis relA deletion strain TW30, and is shown to complement the growth defects of TW30. The recombinant Rel Bst protein was detected by Western immunoanalysis, and synthesizes guanosine‐3′‐diphosphate‐5′‐(tri)diphosphate ((p)ppGpp) after amino acid stress and carbon starvation. These in vivo data, the genetic organization, and the primary structure compared to other RelA/SpoT homologues provide circumstantial evidence that the identified gene encodes the only (p)ppGpp synthetase in B. stearothermophilus , presumed to serve also as (p)ppGpp hydrolase.