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. 1995 Nov;177(21):6276–6281. doi: 10.1128/jb.177.21.6276-6281.1995

Rhizobium NodI and NodJ proteins play a role in the efficiency of secretion of lipochitin oligosaccharides.

H P Spaink 1, A H Wijfjes 1, B J Lugtenberg 1
PMCID: PMC177469  PMID: 7592394

Abstract

Thin-layer chromatographic analysis of extracts of D-[1-14C]glucosamine-labelled rhizobia was used to analyze the effects of nodI, nodJ, and nodT on secretion of lipochitin oligosaccharide (LCO) signal molecules. Secretion was analyzed by comparing quantities of radiolabelled LCOs present in the cellular and spent growth medium fractions. A second rapid and sensitive method was introduced to estimate the secreted LCO fractions by using D-[1-14C]glucosamine-labelled cells grown in medium supplemented with chitinase. At various times after induction of LCO synthesis, the quantity of degradation products of LCOs was compared with the amount of nondegraded LCOs. In wild-type strains of Rhizobium leguminosarum biovars viciae and trifolii the nodI and nodJ genes (but not the nodT gene) strongly enhance the secretion of LCOs during the first 5 h after the induction of LCO synthesis. In LCO-overproducing strains the enhancement of secretion was observed only during the first 3 h after induction. At times later than 5 h after induction, a significant influence of the presence of the nodI and nodJ genes on LCO secretion was detectable neither in the wild type nor in LCO-overproducing strains. By using plasmids in which the nodI and nodJ genes are cloned separately under control of a flavonoid-inducible promoter, it was shown that both genes are needed for a wild-type level of LCO secretion. Therefore, these results demonstrate that nodI and nodJ play a role in determining the efficiency of LCO secretion.

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Selected References

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  1. Atkinson E. M., Palcic M. M., Hindsgaul O., Long S. R. Biosynthesis of Rhizobium meliloti lipooligosaccharide Nod factors: NodA is required for an N-acyltransferase activity. Proc Natl Acad Sci U S A. 1994 Aug 30;91(18):8418–8422. doi: 10.1073/pnas.91.18.8418. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
  3. Bloemberg G. V., Thomas-Oates J. E., Lugtenberg B. J., Spaink H. P. Nodulation protein NodL of Rhizobium leguminosarum O-acetylates lipo-oligosaccharides, chitin fragments and N-acetylglucosamine in vitro. Mol Microbiol. 1994 Feb;11(4):793–804. doi: 10.1111/j.1365-2958.1994.tb00357.x. [DOI] [PubMed] [Google Scholar]
  4. Canter Cremers H., Spaink H. P., Wijfjes A. H., Pees E., Wijffelman C. A., Okker R. J., Lugtenberg B. J. Additional nodulation genes on the Sym plasmid of Rhizobium leguminosarum biovar viciae. Plant Mol Biol. 1989 Aug;13(2):163–174. doi: 10.1007/BF00016135. [DOI] [PubMed] [Google Scholar]
  5. Carlson R. W., Price N. P., Stacey G. The biosynthesis of rhizobial lipo-oligosaccharide nodulation signal molecules. Mol Plant Microbe Interact. 1994 Nov-Dec;7(6):684–695. doi: 10.1094/mpmi-7-0684. [DOI] [PubMed] [Google Scholar]
  6. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Downie J. A. Signalling strategies for nodulation of legumes by rhizobia. Trends Microbiol. 1994 Sep;2(9):318–324. doi: 10.1016/0966-842x(94)90448-0. [DOI] [PubMed] [Google Scholar]
  8. Dénarié J., Cullimore J. Lipo-oligosaccharide nodulation factors: a minireview new class of signaling molecules mediating recognition and morphogenesis. Cell. 1993 Sep 24;74(6):951–954. doi: 10.1016/0092-8674(93)90717-5. [DOI] [PubMed] [Google Scholar]
  9. Geremia R. A., Mergaert P., Geelen D., Van Montagu M., Holsters M. The NodC protein of Azorhizobium caulinodans is an N-acetylglucosaminyltransferase. Proc Natl Acad Sci U S A. 1994 Mar 29;91(7):2669–2673. doi: 10.1073/pnas.91.7.2669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Göttfert M. Regulation and function of rhizobial nodulation genes. FEMS Microbiol Rev. 1993 Jan;10(1-2):39–63. doi: 10.1111/j.1574-6968.1993.tb05863.x. [DOI] [PubMed] [Google Scholar]
  11. Heidstra R., Geurts R., Franssen H., Spaink H. P., Van Kammen A., Bisseling T. Root Hair Deformation Activity of Nodulation Factors and Their Fate on Vicia sativa. Plant Physiol. 1994 Jul;105(3):787–797. doi: 10.1104/pp.105.3.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. John M., Röhrig H., Schmidt J., Wieneke U., Schell J. Rhizobium NodB protein involved in nodulation signal synthesis is a chitooligosaccharide deacetylase. Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):625–629. doi: 10.1073/pnas.90.2.625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. McKay I. A., Djordjevic M. A. Production and Excretion of Nod Metabolites by Rhizobium leguminosarum bv. trifolii Are Disrupted by the Same Environmental Factors That Reduce Nodulation in the Field. Appl Environ Microbiol. 1993 Oct;59(10):3385–3392. doi: 10.1128/aem.59.10.3385-3392.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Orgambide G. G., Lee J., Hollingsworth R. I., Dazzo F. B. Structurally diverse chitolipooligosaccharide nod factors accumulate primarily in membranes of wild type Rhizobium leguminosarum biovar trifolii. Biochemistry. 1995 Mar 21;34(11):3832–3840. doi: 10.1021/bi00011a041. [DOI] [PubMed] [Google Scholar]
  15. Rivilla R., Downie J. A. Identification of a Rhizobium leguminosarum gene homologous to nodT but located outside the symbiotic plasmid. Gene. 1994 Jun 24;144(1):87–91. doi: 10.1016/0378-1119(94)90208-9. [DOI] [PubMed] [Google Scholar]
  16. Röhrig H., Schmidt J., Wieneke U., Kondorosi E., Barlier I., Schell J., John M. Biosynthesis of lipooligosaccharide nodulation factors: Rhizobium NodA protein is involved in N-acylation of the chitooligosaccharide backbone. Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3122–3126. doi: 10.1073/pnas.91.8.3122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schlaman H. R., Okker R. J., Lugtenberg B. J. Subcellular localization of the Rhizobium leguminosarum nodI gene product. J Bacteriol. 1990 Sep;172(9):5486–5489. doi: 10.1128/jb.172.9.5486-5489.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Spaink H. P., Aarts A., Stacey G., Bloemberg G. V., Lugtenberg B. J., Kennedy E. P. Detection and separation of Rhizobium and Bradyrhizobium Nod metabolites using thin-layer chromatography. Mol Plant Microbe Interact. 1992 Jan-Feb;5(1):72–80. doi: 10.1094/mpmi-5-072. [DOI] [PubMed] [Google Scholar]
  19. Spaink H. P., Sheeley D. M., van Brussel A. A., Glushka J., York W. S., Tak T., Geiger O., Kennedy E. P., Reinhold V. N., Lugtenberg B. J. A novel highly unsaturated fatty acid moiety of lipo-oligosaccharide signals determines host specificity of Rhizobium. Nature. 1991 Nov 14;354(6349):125–130. doi: 10.1038/354125a0. [DOI] [PubMed] [Google Scholar]
  20. Spaink H. P., Wijfjes A. H., van der Drift K. M., Haverkamp J., Thomas-Oates J. E., Lugtenberg B. J. Structural identification of metabolites produced by the NodB and NodC proteins of Rhizobium leguminosarum. Mol Microbiol. 1994 Sep;13(5):821–831. doi: 10.1111/j.1365-2958.1994.tb00474.x. [DOI] [PubMed] [Google Scholar]
  21. Surin B. P., Watson J. M., Hamilton W. D., Economou A., Downie J. A. Molecular characterization of the nodulation gene, nodT, from two biovars of Rhizobium leguminosarum. Mol Microbiol. 1990 Feb;4(2):245–252. doi: 10.1111/j.1365-2958.1990.tb00591.x. [DOI] [PubMed] [Google Scholar]
  22. Tsai C. M., Frasch C. E. A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem. 1982 Jan 1;119(1):115–119. doi: 10.1016/0003-2697(82)90673-x. [DOI] [PubMed] [Google Scholar]
  23. Vázquez M., Santana O., Quinto C. The NodL and NodJ proteins from Rhizobium and Bradyrhizobium strains are similar to capsular polysaccharide secretion proteins from gram-negative bacteria. Mol Microbiol. 1993 Apr;8(2):369–377. doi: 10.1111/j.1365-2958.1993.tb01580.x. [DOI] [PubMed] [Google Scholar]
  24. de Maagd R. A., Lugtenberg B. Fractionation of Rhizobium leguminosarum cells into outer membrane, cytoplasmic membrane, periplasmic, and cytoplasmic components. J Bacteriol. 1986 Sep;167(3):1083–1085. doi: 10.1128/jb.167.3.1083-1085.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]

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