Abstract
Monospecific, polyclonal antibodies to the nodC and nodA gene products of Rhizobium meliloti were used in combination with immunogold labeling and transmission electron microscopy to localize the NodC and NodA proteins in cultures of R. meliloti. Both NodC and NodA were detected in the cytoplasm and cell envelope in thin sections of free-living rhizobia treated with luteolin, a known inducer of nod gene expression; however, only NodC was detected on cell surfaces when immunolabeling was performed with intact induced cells. In view of biochemical data characterizing NodC as an outer membrane protein with a large extracellular domain, the pattern of immunolabeling on thin sections suggests that NodC is produced on free cytoplasmic ribosomes prior to assembly in the membrane. The pattern of NodA labeling on thin sections is consistent with biochemical data detecting NodA in both soluble and membrane fractions of NodA-overexpressing strains of R. meliloti.
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Selected References
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- Banfalvi Z., Nieuwkoop A., Schell M., Besl L., Stacey G. Regulation of nod gene expression in Bradyrhizobium japonicum. Mol Gen Genet. 1988 Nov;214(3):420–424. doi: 10.1007/BF00330475. [DOI] [PubMed] [Google Scholar]
- Brewin N. J., Robertson J. G., Wood E. A., Wells B., Larkins A. P., Galfre G., Butcher G. W. Monoclonal antibodies to antigens in the peribacteroid membrane from Rhizobium-induced root nodules of pea cross-react with plasma membranes and Golgi bodies. EMBO J. 1985 Mar;4(3):605–611. doi: 10.1002/j.1460-2075.1985.tb03673.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Egelhoff T. T., Fisher R. F., Jacobs T. W., Mulligan J. T., Long S. R. Nucleotide sequence of Rhizobium meliloti 1021 nodulation genes: nodD is read divergently from nodABC. DNA. 1985 Jun;4(3):241–248. doi: 10.1089/dna.1985.4.241. [DOI] [PubMed] [Google Scholar]
- Fisher R. F., Tu J. K., Long S. R. Conserved Nodulation Genes in Rhizobium meliloti and Rhizobium trifolii. Appl Environ Microbiol. 1985 Jun;49(6):1432–1435. doi: 10.1128/aem.49.6.1432-1435.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fortin M. G., Zelechowska M., Verma D. P. Specific targeting of membrane nodulins to the bacteroid-enclosing compartment in soybean nodules. EMBO J. 1985 Dec 1;4(12):3041–3046. doi: 10.1002/j.1460-2075.1985.tb04043.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fuerst J. A., Perry J. W. Demonstration of lipopolysaccharide on sheathed flagella of Vibrio cholerae O:1 by protein A-gold immunoelectron microscopy. J Bacteriol. 1988 Apr;170(4):1488–1494. doi: 10.1128/jb.170.4.1488-1494.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- John M., Schmidt J., Wieneke U., Kondorosi E., Kondorosi A., Schell J. Expression of the nodulation gene nod C of Rhizobium meliloti in Escherichia coli: role of the nod C gene product in nodulation. EMBO J. 1985 Oct;4(10):2425–2430. doi: 10.1002/j.1460-2075.1985.tb03951.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- John M., Schmidt J., Wieneke U., Krüssmann H. D., Schell J. Transmembrane orientation and receptor-like structure of the Rhizobium meliloti common nodulation protein NodC. EMBO J. 1988 Mar;7(3):583–588. doi: 10.1002/j.1460-2075.1988.tb02850.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marvel D. J., Kuldau G., Hirsch A., Richards E., Torrey J. G., Ausubel F. M. Conservation of nodulation genes between Rhizobium meliloti and a slow-growing Rhizobium strain that nodulates a nonlegume host. Proc Natl Acad Sci U S A. 1985 Sep;82(17):5841–5845. doi: 10.1073/pnas.82.17.5841. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mulligan J. T., Long S. R. Induction of Rhizobium meliloti nodC expression by plant exudate requires nodD. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6609–6613. doi: 10.1073/pnas.82.19.6609. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nguyen T., Zelechowska M., Foster V., Bergmann H., Verma D. P. Primary structure of the soybean nodulin-35 gene encoding uricase II localized in the peroxisomes of uninfected cells of nodules. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5040–5044. doi: 10.1073/pnas.82.15.5040. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nieuwkoop A. J., Banfalvi Z., Deshmane N., Gerhold D., Schell M. G., Sirotkin K. M., Stacey G. A locus encoding host range is linked to the common nodulation genes of Bradyrhizobium japonicum. J Bacteriol. 1987 Jun;169(6):2631–2638. doi: 10.1128/jb.169.6.2631-2638.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Peters N. K., Frost J. W., Long S. R. A plant flavone, luteolin, induces expression of Rhizobium meliloti nodulation genes. Science. 1986 Aug 29;233(4767):977–980. doi: 10.1126/science.3738520. [DOI] [PubMed] [Google Scholar]
- Rossen L., Johnston A. W., Downie J. A. DNA sequence of the Rhizobium leguminosarum nodulation genes nodAB and C required for root hair curling. Nucleic Acids Res. 1984 Dec 21;12(24):9497–9508. doi: 10.1093/nar/12.24.9497. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rossen L., Shearman C. A., Johnston A. W., Downie J. A. The nodD gene of Rhizobium leguminosarum is autoregulatory and in the presence of plant exudate induces the nodA,B,C genes. EMBO J. 1985 Dec 16;4(13A):3369–3373. doi: 10.1002/j.1460-2075.1985.tb04092.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmidt J., John M., Kondorosi E., Kondorosi A., Wieneke U., Schröder G., Schröder J., Schell J. Mapping of the protein-coding regions of Rhizobium meliloti common nodulation genes. EMBO J. 1984 Aug;3(8):1705–1711. doi: 10.1002/j.1460-2075.1984.tb02035.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmidt J., John M., Wieneke U., Krüssmann H. D., Schell J. Expression of the nodulation gene nodA in Rhizobium meliloti and localization of the gene product in the cytosol. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9581–9585. doi: 10.1073/pnas.83.24.9581. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schmidt J., Wingender R., John M., Wieneke U., Schell J. Rhizobium meliloti nodA and nodB genes are involved in generating compounds that stimulate mitosis of plant cells. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8578–8582. doi: 10.1073/pnas.85.22.8578. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tommassen J., Leunissen J., van Damme-Jongsten M., Overduin P. Failure of E. coli K-12 to transport PhoE-LacZ hybrid proteins out of the cytoplasm. EMBO J. 1985 Apr;4(4):1041–1047. doi: 10.1002/j.1460-2075.1985.tb03736.x. [DOI] [PMC free article] [PubMed] [Google Scholar]