1887

Abstract

Acyl carrier proteins (ACPs) are small acidic proteins that carry growing acyl chains during fatty acid or polyketide synthesis. In rhizobia, there are four different and well-characterized ACPs: AcpP, NodF, AcpXL and RkpF. The genome sequence of 1021 reveals two additional ORFs that possibly encode additional ACPs. One of these, , is located on the plasmid pSymB as part of an operon. The genes of the operon encode a putative asparagine synthetase (AsnB), the predicted ACP (SMb20651), a putative long-chain fatty acyl-CoA ligase (SMb20650) and a putative ammonium-dependent NAD synthetase (NadE1). When SMb20651 was overexpressed in [H]-alanine, a biosynthetic building block of 4′-phosphopantetheine, was incorporated into the protein . The purified SMb20651 was modified with 4′-phosphopantetheine in the presence of holo-ACP synthase (AcpS). Also, holo-SMb20651 was modified with a malonyl group by malonyl CoA-ACP transacylase. In , coexpression of SMb20651 together with other proteins such as AcpS and SMb20650 led to the formation of additional forms of SMb20651. In this bacterium, acylation of SMb20651 with C12 : 0 or C18 : 0 fatty acids was detected, demonstrating that this protein is involved in fatty acid biosynthesis or transfer. Expression of SMb20651 was detected in as holo-SMb20651 and acyl-SMb20651.

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2009-01-01
2020-02-22
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References

  1. Altschul S. F., Madden T. L., Schäfer A. A., Zhang J., Zhang Z., Miller W., Lipman D. J.. 1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res25:3389–3402
    [Google Scholar]
  2. Becker A., Bergès H., Krol E., Bruand C., Rüberg S., Capela D., Lauber E., Meilhoc E., Ampe F.. other authors 2004; Global changes in gene expression in Sinorhizobium meliloti 1021 under microoxic and symbiotic conditions. Mol Plant Microbe Interact17:292–303
    [Google Scholar]
  3. Beringer J. E.. 1974; R factor transfer in Rhizobium leguminosarum. J Gen Microbiol84:188–198
    [Google Scholar]
  4. Brozek K. A., Carlson R. W., Raetz C. R.. 1996; A special acyl carrier protein for transferring long hydroxylated fatty acids to lipid A in Rhizobium. J Biol Chem271:32126–32136
    [Google Scholar]
  5. Burris R. H.. 1972; Nitrogen fixation assay-methods and techniques. Methods Enzymol24:415–431
    [Google Scholar]
  6. Byers D. M., Gong H.. 2007; Acyl carrier protein: structure–function relationships in a conserved multifunctional protein family. Biochem Cell Biol85:649–662
    [Google Scholar]
  7. Chao T. C., Buhrmester J., Hansmeier N., Pühler A., Weidner S.. 2005; Role of the regulatory gene rirA in the transcriptional response of Sinorhizobium meliloti to iron limitation. Appl Environ Microbiol71:5969–5982
    [Google Scholar]
  8. Cox R. J., Hitchman T. S., Byrom K. J., Findlow I. S., Tanner J. A., Crosby J., Simpson T. J.. 1997; Post-translational modification of heterologously expressed Streptomyces type II polyketide synthase acyl carrier proteins. FEBS Lett405:267–272
    [Google Scholar]
  9. Ditta G., Schmidhauser T., Yacobson E., Lu P., Liang X. W., Finlay D. R., Helsinki D. R.. 1985; Plasmids related to the broad host range vector, pRK29, useful for gene cloning and monitoring expression. Plasmid 13:149–153
    [Google Scholar]
  10. Domínguez-Ferreras A., Pérez-Arnedo R., Becker A., Olivares J., Soto M. J., Sanjuán J.. 2006; Transcriptome profiling reveals the importance of plasmid pSymB for osmoadaptation of Sinorhizobium meliloti. J Bacteriol188:7617–7625
    [Google Scholar]
  11. Dulley J. R., Grieve P. A.. 1975; A simple technique for eliminating interference by detergents in the Lowry method of protein determination. Anal Biochem64:136–141
    [Google Scholar]
  12. Epple G., Van der Drift K. M. G. M., Thomas-Oates J., Geiger O.. 1998; Characterization of a novel acyl carrier protein, RkpF, encoded by an operon involved in capsular polysaccharide biosynthesis in Sinorhizobium meliloti. J Bacteriol180:4950–4954
    [Google Scholar]
  13. Ferguson G. P., Roop R. M. II, Walker G. C.. 2002; Deficiency of Sinorhizobium meliloti bacA mutant in alfalfa symbiosis correlates with the alteration of cell envelope. J Bacteriol184:5625–5632
    [Google Scholar]
  14. Figurski D. H., Helsinki D. R.. 1979; Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A76:1648–1652
    [Google Scholar]
  15. Finan T. M., Hartwieg E., LeMieux K., Bergman K., Walker G. C., Signer E.. 1984; General transduction in Rhizobium meliloti. J Bacteriol159:120–124
    [Google Scholar]
  16. Flugel R. S., Hwangbo Y., Lambalot R. H., Cronan J. E., Walsh C. T.. 2000; Holo-(acyl carrier protein) synthase and phosphopantetheinyl transfer in Escherichia coli. J Biol Chem275:959–978
    [Google Scholar]
  17. Fuqua C., Greenberg E. P.. 2002; Listening in on bacteria: acyl-homoserine lactone signalling. Nat Rev Mol Cell Biol3:685–695
    [Google Scholar]
  18. Galibert F., Finan T. M., Long S. R., Puhler A., Abola P., Ampe F., Barloy-Hubler F., Barnett M. J., Becker A.. other authors 2001; The composite genome of the legume symbiont Sinorhizobium meliloti. Science293:668–672
    [Google Scholar]
  19. Geiger O., López-Lara I. M.. 2002; Rhizobial acyl carrier proteins and their roles in the formation of bacterial cell surface components that are required for the development of nitrogen-fixing root nodules on legume hosts. FEMS Microbiol Lett208:153–162
    [Google Scholar]
  20. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L.. 1989; Site directed mutagenesis by overlap extension using polymerase chain reaction. Gene77:51–59
    [Google Scholar]
  21. Jackowski S., Rock C. O.. 1983; Ratio of active to inactive forms of acyl carrier protein in Escherichia coli. J Biol Chem258:15186–15191
    [Google Scholar]
  22. Jiang Y., Chan C. H., Cronan J. E.. 2006; The soluble acyl-acyl carrier protein synthetase of Vibrio harveyi B392 is a member of the medium chain acyl-CoA synthetase family. Biochemistry45:10008–10019
    [Google Scholar]
  23. Kato J. Y., Funa N., Watanabe H., Ohnishi Y., Horinouchi S.. 2007; Biosynthesis of γ-butyrolactone autoregulators that switch on secondary metabolism and morphological development in Streptomyces. Proc Natl Acad Sci U S A104:2378–2383
    [Google Scholar]
  24. Kovach M. E., Elzer P. H., Hill D. S., Robertson G. T., Farris M. A., Roop R. M. II, Peterson K. M.. 1995; Four new derivatives of the broad-host range vector pBBR1MCS, carrying different antibiotic resistance cassettes. Gene166:175–176
    [Google Scholar]
  25. Lai J. R., Koglin A., Walsh C. T.. 2006; Carrier protein structure and recognition in polyketide and nonribosomal peptide biosynthesis. Biochemistry45:14869–14879
    [Google Scholar]
  26. Lambalot R. H., Gehring A. M., Flugel R. S., Zuber P., LaCelle M., Marahiel M. A., Reid R., Khosla C., Walsh C. T.. 1996; A new enzyme superfamily: the phosphopantetheinyl transferases. Chem Biol3:923–936
    [Google Scholar]
  27. López-Lara I. M., Geiger O.. 2000; Expression and purification of four different acyl carrier proteins. Microbiology146:839–849
    [Google Scholar]
  28. López-Lara I. M., Geiger O.. 2001; The nodulation protein NodG shows the enzymatic activity of an 3-oxoacyl-acyl carrier protein reductase. Mol Plant Microbe Interact14:349–357
    [Google Scholar]
  29. Miller J. H.. 1972; Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  30. Neuhaus F. C., Baddiley J.. 2003; A continuum of anionic charge: structure, and functions of d-alanyl-teichoic acids in Gram-positive bacteria. Microbiol Mol Biol Rev67:686–723
    [Google Scholar]
  31. Olivares J., Casadesus J., Bedmar E. J.. 1980; Method for testing degree of infectivity of Rhizobium meliloti strains. Appl Environ Microbiol39:967–970
    [Google Scholar]
  32. Østeras M., Boncopagni E., Vincent N., Poggi M. C., Le Rudulier D.. 1998; Presence of a gene encoding choline sulfatase in Sinorhizobium meliloti bet operon: choline- O-sulfate is metabolised into glycine betaine. Proc Natl Acad Sci U S A95:11394–11399
    [Google Scholar]
  33. Platt M. W., Miller K. J., Lane W. S., Kennedy E. P.. 1990; Isolation and characterization of the constitutive acyl carrier protein from Rhizobium meliloti. J Bacteriol172:5440–5444
    [Google Scholar]
  34. Post-Beittenmiller D., Jaworski J. G., Ohlrogge J. B.. 1991; In vivo pools of free and acylated acyl carrier proteins in spinach. Evidence for sites of regulation of fatty acid biosynthesis. J Biol Chem266:1858–1865
    [Google Scholar]
  35. Ritsema T., Gehring A. M., Stuitje A. R., van der Drift K. M., Dandal I., Lambalot R. H., Walsh C. T., Thomas-Oates J. E., Lugtenberg B. J., Spaink H. P.. 1998; Functional analysis of an interspecies chimera of acyl carrier proteins indicates a specialized domain for protein recognition. Mol Gen Genet257:641–648
    [Google Scholar]
  36. Robertsen B. K., Aitman P., Darvill A. G., McNeil M., Alberstein P.. 1981; The structure of acidic extracellular polysaccharides secreted by Rhizobium leguminosarum and Rhizobium trifolii. Plant Physiol67:389–400
    [Google Scholar]
  37. Sambrook J., Russell D. R.. 2001; Molecular Cloning: a Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  38. Schäfer A., Tauch A., Jäger W., Kalinowski J., Thierbach G., Pühler A.. 1994; Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene145:69–73
    [Google Scholar]
  39. Sherwood M. T.. 1970; Improved synthetic medium for the growth of Rhizobium. J Appl Bacteriol33:708–713
    [Google Scholar]
  40. Sohlenkamp C., de Rudder K. E., Geiger O.. 2004; Phosphatidylethanolamine is not essential for growth of Sinorhizobium meliloti on complex culture media. J Bacteriol186:1667–1677
    [Google Scholar]
  41. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W.. 1990; Use of a T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol185:60–89
    [Google Scholar]
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