Membrane glycerophospholipid biosynthesis in Neisseria meningitidis and Neisseria gonorrhoeae: identification, characterization, and mutagenesis of a lysophosphatidic acid acyltransferase.
Lysophosphatidic acid (LPA) acyltransferases of Neisseria meningitidis and Neisseria gonorrhoeae were identified which share homology with other prokaryotic and eukaryotic LPA acyltransferases. In Escherichia coli, the conversion of LPA to phosphatidic acid, performed by the 1-acyl-sn-glycerol-3-phosphate acyltransferase PlsC, is a critical intermediate step in the biosynthesis of membrane glycerophospholipids. A Tn916-generated mutant of a serogroup B meningococcal strain was identified that exhibited increased amounts of capsular polysaccharide, as shown by colony immunoblots, and a threefold increase in the number of assembled pili. The single, truncated 3.8 kb Tn916 insertion in the meningococcal mutant was localized within a 771 bp open reading frame, The gonococcal equivalent of this gene was identified by transformation with the cloned meningococcal mutant gene. In N. gonorrhoeae, the mutation increased piliation fivefold. The insertions were found to be within a gene that was subsequently designated nlaA (neisserial LPA acyltransferase). The predicted neisserial LPA acyltransferases were homologous (>20% identity, >40% amino acid similarity) to the family of PlsC protein homologues. A cloned copy of the meningococcal nlaA gene complemented in trans a temperature-sensitive E. coli PlsCts- mutant. Tn916 and omega-cassette insertional inactivations of the neisserial nlaA genes altered the membrane glycerophospholipid compositions of both N. meningitidis and N. gonorrhoeae but were not lethal. Therefore, the pathogenic Neisseria spp. appear to be able to utilize alternative enzyme(s) to produce phosphatidic acid. This hypothesis is supported by the observation that, although the amounts of mature glycerophospholipids were altered in the meningococcal and the gonococcal nlaA mutants, glycerophospholipid synthesis was detectable at significant levels. In addition, acyltransferase enzymatic activity, while reduced in the gonococcal nlaA mutant, was increased in the meningococcal nlaA mutant. We postulate that the pathogenic Neisseria spp. are able to utilize alternate acyltransferases to produce glycerophospholipids in the absence of nlaA enzymatic activity. Implementation of these secondary enzymes results in alterations of glycerophospholipid composition that lead to pleiotropic effects on the cell surface components, including effects on capsule and piliation.
Swartley
J S
JS
Emory University School of Medicine and VA Medical Center, Atlanta, Georgia 30033, USA.
Balthazar
J T
JT
Coleman
J
J
Shafer
W M
WM
Stephens
D S
DS
D000217:Acyltransferases / Q000235:genetics* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D016667:Bacterial Capsules / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D004926:Escherichia coli / Q000235:genetics; D010861:Fimbriae, Bacterial / Q000378:metabolism / Q000648:ultrastructure; D015870:Gene Expression; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009154:Mutation; D009344:Neisseria gonorrhoeae / Q000737:chemistry / Q000201:enzymology* / Q000472:pathogenicity; D009345:Neisseria meningitidis / Q000737:chemistry / Q000201:enzymology* / Q000472:pathogenicity; D010957:Plasmids; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D000217:Acyltransferases; C018764:2-acylglycerophosphate acyltransferase