RRC ID 48565
著者 Matsumoto K, Okada M, Horikoshi Y, Matsuzaki H, Kishi T, Itaya M, Shibuya I.
タイトル Cloning, sequencing, and disruption of the Bacillus subtilis psd gene coding for phosphatidylserine decarboxylase.
ジャーナル J Bacteriol
Abstract The psd gene of Bacillus subtilis Marburg, encoding phosphatidylserine decarboxylase, has been cloned and sequenced. It encodes a polypeptide of 263 amino acid residues (deduced molecular weight of 29,689) and is located just downstream of pss, the structural gene for phosphatidylserine synthase that catalyzes the preceding reaction in phosphatidylethanolamine synthesis (M. Okada, H. Matsuzaki, I. Shibuya, and K. Matsumoto, J. Bacteriol. 176:7456-7461, 1994). Introduction of a plasmid containing the psd gene into temperature-sensitive Escherichia coli psd-2 mutant cells allowed growth at otherwise restrictive temperature. Phosphatidylserine was not detected in the psd-2 mutant cells harboring the plasmid; it accumulated in the mutant up to 29% of the total phospholipids without the plasmid. An enzyme activity that catalyzes decarboxylation of 14C-labeled phosphatidylserine to form phosphatidylethanolamine was detected in E. coli psd-2 cells harboring a Bacillus psd plasmid. E. coli cells harboring the psd plasmid, the expression of which was under the control of the T7phi10 promoter, produced proteins of 32 and 29 kDa upon induction. A pulse-labeling experiment suggested that the 32-kDa protein is the primary translation product and is processed into the 29-kDa protein. The psd gene, together with pss, was located by Southern hybridization to the 238- to 306-kb SfiI-NotI fragment of the chromosome. A B. subtilis strain harboring an interrupted psd allele, psd1::neo, was constructed. The null psd mutant contained no phosphatidylethanolamine and accumulated phosphatidylserine. It grew well without supplementation of divalent cations which are essential for the E. coli pssA null mutant lacking phosphatidylethanolamine. In both the B. subtilis null pss and psd mutants, glucosyldiacylglycerol content increased two- to fourfold. The results suggest that the lack of phosphatidylethanolamine in the B. subtilis membrane may be compensated for by the increases in the contents of glucosyldiacylglycerols by an unknown mechanism.
巻・号 180(1)
ページ 100-6
公開日 1998-1-1
DOI 10.1128/JB.180.1.100-106.1998
PMID 9422599
PMC PMC106855
MeSH Amino Acid Sequence Bacillus subtilis / enzymology Bacillus subtilis / genetics* Base Sequence CDPdiacylglycerol-Serine O-Phosphatidyltransferase / genetics Carboxy-Lyases / chemistry Carboxy-Lyases / genetics* Carboxy-Lyases / metabolism Cloning, Molecular Escherichia coli / genetics Genes, Bacterial / genetics* Genetic Complementation Test Molecular Sequence Data Molecular Weight Phospholipids / analysis Protein Processing, Post-Translational Restriction Mapping Sequence Analysis, DNA Sequence Homology, Amino Acid Temperature
IF 3.006
引用数 41
WOS 分野 MICROBIOLOGY
リソース情報
原核生物(枯草菌) MBS424 MBS425 MBS426