Involvement of PlsX and the acyl-phosphate dependent sn-glycerol-3-phosphate acyltransferase PlsY in the initial stage of glycerolipid synthesis in Bacillus subtilis.
The gene responsible for the first acylation of sn-glycerol-3-phosphate (G3P) in Bacillus subtilis has not yet been determined with certainty. The product of this first acylation, lysophosphatidic acid (LPA), is subsequently acylated again to form phosphatidic acid (PA), the primary precursor to membrane glycerolipids. A novel G3P acyltransferase (GPAT), the gene product of plsY, which uses acyl-phosphate formed by the plsX gene product, has recently been found to synthesize LPA in Streptococcus pneumoniae. We found that in B. subtilis growth arrests after repression of either a plsY homologue or a plsX homologue were overcome by expression of E. coli plsB, which encodes an acyl-acylcarrier protein (acyl-ACP)-dependent GPAT, although in the case of plsX repression a high level of plsB expression was required. B. subtilis has, therefore, a capability to use the acyl-ACP dependent GPAT of PlsB. Simultaneous expression of plsY and plsX suppressed the glycerol requirement of a strict glycerol auxotrophic derivative of the E. coli plsB26 mutant, although either one alone did not. Membrane fractions from B. subtilis cells catalyzed palmitoylphosphate-dependent acylation of [14C]-labeled G3P to synthesize [14C]-labeled LPA, whereas those from DeltaplsY cells did not. The results indicate unequivocally that PlsY is an acyl-phosphate dependent GPAT. Expression of plsX corrected the glycerol auxotrophy of a DeltaygiH (the deleted allele of an E. coli homologue of plsY) derivative of BB26-36 (plsB26 plsX50), suggesting an essential role of plsX other than substrate supply for acyl-phosphate dependent LPA synthesis. Two-hybrid examinations suggested that PlsY is associated with PlsX and that each may exist in multimeric form.
Hara
Yoshinori
Y
Department of Biochemistry and Molecular Biology, Graduate School of Science and Engineering, Saitama University.
Seki
Masahide
M
Matsuoka
Satoshi
S
Hara
Hiroshi
H
Yamashita
Atsushi
A
Matsumoto
Kouji
K
D000123:Acetyltransferases / Q000378:metabolism; D000215:Acylation; D001412:Bacillus subtilis / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D002250:Carbon Radioisotopes; D002462:Cell Membrane / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000378:metabolism*; D005992:Glycerol-3-Phosphate O-Acyltransferase / Q000378:metabolism*; D020404:Glycerophospholipids / Q000096:biosynthesis*; D010957:Plasmids / Q000235:genetics; D020798:Two-Hybrid System Techniques
D001426:Bacterial Proteins; D002250:Carbon Radioisotopes; D017931:DNA Primers; D020404:Glycerophospholipids; C088150:plsX protein, bacteria; D000123:Acetyltransferases; C512507:PlsB protein, E coli; D005992:Glycerol-3-Phosphate O-Acyltransferase