Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Apr;179(7):2132–2140. doi: 10.1128/jb.179.7.2132-2140.1997

The rkpGHI and -J genes are involved in capsular polysaccharide production by Rhizobium meliloti.

E Kiss 1, B L Reuhs 1, J S Kim 1, A Kereszt 1, G Petrovics 1, P Putnoky 1, I Dusha 1, R W Carlson 1, A Kondorosi 1
PMCID: PMC178947  PMID: 9079896

Abstract

The first complementation unit of the fix-23 region of Rhizobium meliloti, which comprises six genes (rkpAB-CDEF) exhibiting similarity to fatty acid synthase genes, is required for the production of a novel type of capsular polysaccharide that is involved in root nodule development and structurally analogous to group II K antigens found in Escherichia coli (G. Petrovics, P. Putnoky, R. Reuhs, J. Kim, T. A. Thorp, K. D. Noel, R. W. Carlson, and A. Kondorosi, Mol. Microbiol. 8:1083-1094, 1993; B. L. Reuhs, R. W. Carlson, and J. S. Kim, J. Bacteriol. 175:3570-3580, 1993). Here we present the nucleotide sequence for the other three complementation units of the fix-23 locus, revealing the presence of four additional open reading frames assigned to genes rkpGHI and -J. The putative RkpG protein shares similarity with acyltransferases, RkpH is homologous to short-chain alcohol dehydrogenases, and RkpJ shows significant sequence identity with bacterial polysaccharide transport proteins, such as KpsS of E. coli. No significant homology was found for RkpI. Biochemical and immunological analysis of Tn5 derivatives for each gene demonstrated partial or complete loss of capsular polysaccharides from the cell surface; on this basis, we suggest that all genes in the fix-23 region are required for K-antigen synthesis or transport.

Full Text

The Full Text of this article is available as a PDF (1.5 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Argos P., Rao J. K. Prediction of protein structure. Methods Enzymol. 1986;130:185–207. doi: 10.1016/0076-6879(86)30012-0. [DOI] [PubMed] [Google Scholar]
  3. Boulnois G. J., Roberts I. S., Hodge R., Hardy K. R., Jann K. B., Timmis K. N. Analysis of the K1 capsule biosynthesis genes of Escherichia coli: definition of three functional regions for capsule production. Mol Gen Genet. 1987 Jun;208(1-2):242–246. doi: 10.1007/BF00330449. [DOI] [PubMed] [Google Scholar]
  4. Bronner D., Sieberth V., Pazzani C., Roberts I. S., Boulnois G. J., Jann B., Jann K. Expression of the capsular K5 polysaccharide of Escherichia coli: biochemical and electron microscopic analyses of mutants with defects in region 1 of the K5 gene cluster. J Bacteriol. 1993 Sep;175(18):5984–5992. doi: 10.1128/jb.175.18.5984-5992.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bánfalvi Z., Sakanyan V., Koncz C., Kiss A., Dusha I., Kondorosi A. Location of nodulation and nitrogen fixation genes on a high molecular weight plasmid of R. meliloti. Mol Gen Genet. 1981;184(2):318–325. doi: 10.1007/BF00272925. [DOI] [PubMed] [Google Scholar]
  6. Cross A. S. The biologic significance of bacterial encapsulation. Curr Top Microbiol Immunol. 1990;150:87–95. doi: 10.1007/978-3-642-74694-9_5. [DOI] [PubMed] [Google Scholar]
  7. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Franssen H. J., Vijn I., Yang W. C., Bisseling T. Developmental aspects of the Rhizobium-legume symbiosis. Plant Mol Biol. 1992 May;19(1):89–107. doi: 10.1007/BF00015608. [DOI] [PubMed] [Google Scholar]
  9. Frosch M., Edwards U., Bousset K., Krausse B., Weisgerber C. Evidence for a common molecular origin of the capsule gene loci in gram-negative bacteria expressing group II capsular polysaccharides. Mol Microbiol. 1991 May;5(5):1251–1263. doi: 10.1111/j.1365-2958.1991.tb01899.x. [DOI] [PubMed] [Google Scholar]
  10. Frosch M., Müller A. Phospholipid substitution of capsular polysaccharides and mechanisms of capsule formation in Neisseria meningitidis. Mol Microbiol. 1993 May;8(3):483–493. doi: 10.1111/j.1365-2958.1993.tb01592.x. [DOI] [PubMed] [Google Scholar]
  11. Fuhrmann J., Wollum A. G. Simplified Enzyme-Linked Immunosorbent Assay for Routine Identification of Rhizobium japonicum Antigens. Appl Environ Microbiol. 1985 Apr;49(4):1010–1013. doi: 10.1128/aem.49.4.1010-1013.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Jörnvall H., von Bahr-Lindström H., Jeffery J. Extensive variations and basic features in the alcohol dehydrogenase-sorbitol dehydrogenase family. Eur J Biochem. 1984 Apr 2;140(1):17–23. doi: 10.1111/j.1432-1033.1984.tb08061.x. [DOI] [PubMed] [Google Scholar]
  13. Kannenberg E. L., Brewin N. J. Host-plant invasion by Rhizobium: the role of cell-surface components. Trends Microbiol. 1994 Aug;2(8):277–283. doi: 10.1016/0966-842x(94)90004-3. [DOI] [PubMed] [Google Scholar]
  14. Kroll J. S., Zamze S., Loynds B., Moxon E. R. Common organization of chromosomal loci for production of different capsular polysaccharides in Haemophilus influenzae. J Bacteriol. 1989 Jun;171(6):3343–3347. doi: 10.1128/jb.171.6.3343-3347.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Leigh J. A., Coplin D. L. Exopolysaccharides in plant-bacterial interactions. Annu Rev Microbiol. 1992;46:307–346. doi: 10.1146/annurev.mi.46.100192.001515. [DOI] [PubMed] [Google Scholar]
  16. Pazzani C., Rosenow C., Boulnois G. J., Bronner D., Jann K., Roberts I. S. Molecular analysis of region 1 of the Escherichia coli K5 antigen gene cluster: a region encoding proteins involved in cell surface expression of capsular polysaccharide. J Bacteriol. 1993 Sep;175(18):5978–5983. doi: 10.1128/jb.175.18.5978-5983.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Persson B., Krook M., Jörnvall H. Characteristics of short-chain alcohol dehydrogenases and related enzymes. Eur J Biochem. 1991 Sep 1;200(2):537–543. doi: 10.1111/j.1432-1033.1991.tb16215.x. [DOI] [PubMed] [Google Scholar]
  18. Petrovics G., Putnoky P., Reuhs B., Kim J., Thorp T. A., Noel K. D., Carlson R. W., Kondorosi A. The presence of a novel type of surface polysaccharide in Rhizobium meliloti requires a new fatty acid synthase-like gene cluster involved in symbiotic nodule development. Mol Microbiol. 1993 Jun;8(6):1083–1094. doi: 10.1111/j.1365-2958.1993.tb01653.x. [DOI] [PubMed] [Google Scholar]
  19. Putnoky P., Kondorosi A. Two gene clusters of Rhizobium meliloti code for early essential nodulation functions and a third influences nodulation efficiency. J Bacteriol. 1986 Sep;167(3):881–887. doi: 10.1128/jb.167.3.881-887.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Putnoky P., Petrovics G., Kereszt A., Grosskopf E., Ha D. T., Bánfalvi Z., Kondorosi A. Rhizobium meliloti lipopolysaccharide and exopolysaccharide can have the same function in the plant-bacterium interaction. J Bacteriol. 1990 Sep;172(9):5450–5458. doi: 10.1128/jb.172.9.5450-5458.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Reuhs B. L., Carlson R. W., Kim J. S. Rhizobium fredii and Rhizobium meliloti produce 3-deoxy-D-manno-2-octulosonic acid-containing polysaccharides that are structurally analogous to group II K antigens (capsular polysaccharides) found in Escherichia coli. J Bacteriol. 1993 Jun;175(11):3570–3580. doi: 10.1128/jb.175.11.3570-3580.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Reuhs B. L., Williams M. N., Kim J. S., Carlson R. W., Côté F. Suppression of the Fix- phenotype of Rhizobium meliloti exoB mutants by lpsZ is correlated to a modified expression of the K polysaccharide. J Bacteriol. 1995 Aug;177(15):4289–4296. doi: 10.1128/jb.177.15.4289-4296.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rimokh R., Gadoux M., Berthéas M. F., Berger F., Garoscio M., Deléage G., Germain D., Magaud J. P. FVT-1, a novel human transcription unit affected by variant translocation t(2;18)(p11;q21) of follicular lymphoma. Blood. 1993 Jan 1;81(1):136–142. [PubMed] [Google Scholar]
  24. Roberts I. S., Mountford R., Hodge R., Jann K. B., Boulnois G. J. Common organization of gene clusters for production of different capsular polysaccharides (K antigens) in Escherichia coli. J Bacteriol. 1988 Mar;170(3):1305–1310. doi: 10.1128/jb.170.3.1305-1310.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Williams M. N., Hollingsworth R. I., Klein S., Signer E. R. The symbiotic defect of Rhizobium meliloti exopolysaccharide mutants is suppressed by lpsZ+, a gene involved in lipopolysaccharide biosynthesis. J Bacteriol. 1990 May;172(5):2622–2632. doi: 10.1128/jb.172.5.2622-2632.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES