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Characterization of the Escherichia coli gene for 1-acyl-sn-glycerol-3-phosphate acyltransferase (pIsC)

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Summary

An Escherichia coli strain deficient in 1-acyl-sn-glycerol-3-phosphate acyltransferase activity has previously been isolated, and the gene (plsC) has been shown to map near min 65 on the chromosome. I precisely mapped the location of plsC on the chromosome, and determined its DNA sequence. plsC is located between parC and sufI, and is separated from sufI by 74 bp. Upstream of plsC is parC, separated by 233 bp, which includes an active promoter. parC, plsC, and sufI are all transcribed in the counterclockwise direction on the chromosome, possibly in an operon with multiple promoters. The amino-terminal sequence of the partially purified protein, combined with the DNA sequence, reveal 1-acyl-sn-glycerol-3-phosphate acyltransferase to be a 27.5 kDa highly basic protein. The plsC gene product, 1-acyl-sn-glycerol-3-phosphate acyltransferase, is localized to the cytoplasmic membrane of the cell. The amino-terminal sequence of the purified protein reveals the first amino acid to be a blocked methionine residue, most probably a formyl-methionine. The amino acid sequence of 1-acyl-sn-glycerol-3-phosphate acyltransferase has a short region of homology to two other E. coli acyltransferases that utilize acyl-acyl carrier protein as the acyl donor, sn-glycerol-3-phosphate acyltransferase and UDP-N-acetyl-glucosamine acyltransferase (involved in lipid A biosynthesis).

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References

  • Anderson CW, Baum PR, Gesteland RF (1973) Processing of adenovirus 2 induced proteins. J Virol 12:241–252

    Google Scholar 

  • Belfaiza J, Parsot C, Martel A, Bouthier de la Tour C, Margarita D, Cohen GN, Saint-Girons I (1986) Evolution in biosynthetic pathways: Two enzymes catalyzing consecutive steps in methionine biosynthesis originate from a common ancestor and possess a similar regulatory region. Proc Natl Acad Sci USA 83:867–871

    Google Scholar 

  • Boom TV, Cronan JE Jr (1989) Genetics and regulation of bacterial lipid metabolism. Annu Rev Microbiol 43:317–343

    Google Scholar 

  • Bullock WO, Fernandez JM, Short JM (1987) XLl-Blue: a high efficiency plasmid transforming recA Escherichia coli strain with beta-galactosidase selection. BioTechniques 5:376–379

    Google Scholar 

  • Cao Y, Oo K-C, Huang AHC (1990) Lysophosphatidate acyltransferase in microsomes from maturing seeds of meadowfoam (Limnanthes alba). Plant Physiol 94:1199–1206

    Google Scholar 

  • Chou PY, Fasman GD (1978) Empirical predictions of protein conformation. Annu Rev Biochem 47:251–276

    Google Scholar 

  • Clarke L, Carbon J (1976) A colony bank containing synthetic Col EI hybrid plasmids representative of the entire E. coli genome. Cell 9:91–99

    Google Scholar 

  • Coleman J (1990) Characterization of Escherichia coli cells deficient in 1-acyl-sn-glycerol-3-phosphate acyltransferase activity. J Biol Chem 265:17215–17221

    Google Scholar 

  • Coleman J, Raetz CRH (1988) The first committed step of lipid A biosynthesis in Escherichia coli: Sequence of the lpxA gene. J Bacteriol 170:1268–1274

    Google Scholar 

  • Cronan JE Jr, Rock CO (1987) Biosynthesis of membrane lipids. In: Neidhardt FC (ed) Escherichia coli and Salmonella typhimurium: cellular and molecular biology, vol 1. American Society for Microbiology, Washington, DC, pp 474–497

    Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395

    Google Scholar 

  • Filip C, Fletcher G, Wulff JL, Earhart CF (1973) Solubilization of the cytoplasmic membrane of Escherichia coli by the ionic detergent sodium-lauryl sarcosinate. J Bacteriol 115:717–722

    Google Scholar 

  • Friedman DI (1988) Integration Host Factor: a protein for all reasons. Cell 55:545–554

    Google Scholar 

  • Fuller RS, Funell BE, Kornberg A (1984) The dnaA protein complex with the Escherichia coli chromosomal replication origin (oriC) and other DNA sites. Cell 38:889–900

    Google Scholar 

  • Gribskov M, Devereux J, Burgess RR (1984) The codon preference plot: graphic analysis of protein coding sequences and prediction of gene expression. Nucleic Acids Res 12:539–549

    Google Scholar 

  • Hirel P-H, Schmitter J-M, Dessen P, Fayat G, Blanquet S (1989) Extent of N-terminal methionine excision from Escherichia coli proteins is governed by the side-chain length of the penultimate amino acid. Proc Natl Acad Sci USA 86:8247–8251

    Google Scholar 

  • Jovin TM (1973a) Multiphasic zone electrophoresis. 3. Further analysis and new forms of discontinuous buffer systems. Biochemistry 12:890–898

    Google Scholar 

  • Jovin TM (1973b) Multiphasic zone electrophoresis. II. Design of integrated discontinuous buffer systems for analytical and preparative fractionation. Biochemistry 12:879–890

    Google Scholar 

  • Jovin TM (1973c) Multiphasic zone electrophoresis. I. Steady-state moving-boundary systems formed by different electrolyte combinations. Biochemistry 12:871–879

    Google Scholar 

  • Kato J-I, Nishimura Y, Yamada M, Suzuki H, Hirota Y (1988) Gene organization in the region containing a new gene involved in chromosome partitioning in Escherichia coli. J Bacteriol 170:3967–3977

    Google Scholar 

  • Kato J-I, Nishimura Y, Imamura R, Niki H, Hiraga S, Suzuki H (1990) New topoisomerase essential for chromosome segregation in E. coli. Cell 63:393–404

    Google Scholar 

  • Kessels JMM, Ousen H, Van Den Bosch H (1983) Facilitated utilization of endogenously synthesized lysophosphatidic acid by 1-acylglycerophosphate acyltransferase from Escherichia coli. Biochim Biophys Acta 753:227–235

    Google Scholar 

  • Kohara Y, Akiotaka K, Isono K (1987) The physical map of the whole E. coli chromosome: Application of a new strategy for rapid analysis and sorting of a large genomic library. Cell 50:495–508

    Google Scholar 

  • Kozak M (1983) Comparison of initiation of protein synthesis in procaryotes, eucaryotes, and organelles. Microbiol Rev 47:1–45

    Google Scholar 

  • Kyle J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Lightner VA, Bell RM, Modrich P (1983) The DNA sequences encoding plsB and dgk loci of Escherichia coli. J Biol Chem 258:10856–10861

    Google Scholar 

  • Lipman DJ, Pearson WR (1985) Rapid and sensitive protein similarity searches. Science 227:1435–1441

    Google Scholar 

  • Little JW, Mount DW (1982) The SOS regulatory system of Escherichia coli. Cell 29:11–22

    Google Scholar 

  • Luttinger AL, Springer AL, Schmid MB (1991) A cluster of genes that affects nucleoid segregation in Salmonella typhimurium. New Biologist 3:687–697

    Google Scholar 

  • MilliganDL, Koshland DE Jr (1990) The amino terminus of the aspartate chemoreceptors is formylmethionine. J Biol Chem 265:4455–4460

    Google Scholar 

  • Moos M Jr, Nguyen NY, Liu T-Y (1988) Reproducible high yield sequencing of proteins electrophoretically separated and transferred to an inert support. J Biol Chem 263:6005–6008

    Google Scholar 

  • O'Neill MC, Chiafari F (1989) Escherichia coli promoters: A spacing class-dependent search protocol. J Biol Chem 264:5531–5534

    Google Scholar 

  • Olson ER, Flamm EL, Friedman DI (1982) Analysis of nut-R: A region of phage lambda required for antitermination of transcription. Cell 31:61–70

    Google Scholar 

  • Raetz CRH, Dowhan W (1990) Biosynthesis and function of phospholipids in Escherichia coli. J Biol Chem 265:1235–1238

    Google Scholar 

  • Rock CO, Goelz SE, Cronan JE Jr (1981) Phospholipid synthesis in Escherichia coli: Characteristics of fatty acid transfer from acyl-acyl carrier protein to sn-glycerol-3-phosphate. J Biol Chem 256:736–742

    Google Scholar 

  • Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 13:319–353

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • Schmid MB (1990) A locus affecting nucleoid segregation in Salmonella typhimurium. J Bacteriol 172:5416–5424

    Google Scholar 

  • Sheehan JC, Yang D-DH (1957) The use of N-formyl amino acids in peptide synthesis. J Am Chem Soc 80:1154–1158

    Google Scholar 

  • Shine J, Dalgarno L (1974) The 3′ terminal sequence of Escherichia coli 16S ribosome RNA: complementary to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346

    Google Scholar 

  • Snider MD, Kennedy EP (1977) Partial purification of glycerophosphate acyltransferase from Escherichia coli. J Bacteriol 130:1072–1083

    Google Scholar 

  • Stern MJ, Ames GF-L, Smith NH, Robinson EC, Higgins CF (1984) Repetitive extragenic palindromic sequences: a major component of the bacterial genome. Cell 37:1015–1026

    Google Scholar 

  • Sun C, Cao Y, Huang ACH (1988) Acyl Coenzyme A preference of the glycerol phosphate pathway in the microsomes from the maturing seeds of palm, maize, and rapeseed. Plant Physiol 88:56–60

    Google Scholar 

Download references

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  1. Department of Biochemistry and Molecular Biology, Louisiana State University - Medical Center, 1901 Perdido St., 70112, New Orleans, LA, USA

    Jack Coleman

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  1. Jack Coleman
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Communicated by H. Hennecke

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Coleman, J. Characterization of the Escherichia coli gene for 1-acyl-sn-glycerol-3-phosphate acyltransferase (pIsC). Molec. Gen. Genet. 232, 295–303 (1992). https://doi.org/10.1007/BF00280009

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  • DOI: https://doi.org/10.1007/BF00280009

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