• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Feb 1996; 178(3): 745–752.
PMCID: PMC177721

Genetic analysis of Rhizobium meliloti bacA-phoA fusion results in identification of degP: two loci required for symbiosis are closely linked to degP.

Abstract

The function of the Rhizobium meliloti bacA gene, which is a homolog of the Escherichia coli sbmA gene, is required for an intermediate step in nodule development. A strain carrying the bacA386::TnphoA fusion was mutagenized with N-methyl-N'-nitro-N-nitrosoguanidine, and three mutants that had higher levels of alkaline phosphatase activity were identified. The mutations in these strains were recessive and mapped to the same genetic locus. The gene affected by these mutations was identified and sequenced and was found to be a homolog of the E. coli degP gene, which encodes a periplasmic endopeptidase. Although degP function is important for the virulence of certain intracellular pathogens of mammals, it is not required for the R. meliloti-alfalfa symbiosis. The genetic analyses involving degP were complicated by the presence of a locus immediately upstream of depP that was lethal when present in multiple copies in a DegP- background. R. meliloti derivatives carrying insertion mutations in this locus displayed an N,N,N',N'-tetramethyl-p-phenylenediamine oxidase-negative phenotype, elicited the formation of white cylindrical nodules that did not fix nitrogen, and grew slowly in rich medium, suggesting that the locus was a cyc gene encoding a protein involved in the biosynthesis of a component or components of a respiratory chain. The previously identified fix-382::TnphoA, which similarly causes the formation of white cylindrical nodules that do not fix nitrogen, was shown to affect a gene that is separate from this cyc gene but extremely closely linked to it.

Full Text

The Full Text of this article is available as a PDF (278K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. [PubMed]
  • Barsomian GD, Urzainqui A, Lohman K, Walker GC. Rhizobium meliloti mutants unable to synthesize anthranilate display a novel symbiotic phenotype. J Bacteriol. 1992 Jul;174(13):4416–4426. [PMC free article] [PubMed]
  • Bäumler AJ, Kusters JG, Stojiljkovic I, Heffron F. Salmonella typhimurium loci involved in survival within macrophages. Infect Immun. 1994 May;62(5):1623–1630. [PMC free article] [PubMed]
  • Bott M, Bolliger M, Hennecke H. Genetic analysis of the cytochrome c-aa3 branch of the Bradyrhizobium japonicum respiratory chain. Mol Microbiol. 1990 Dec;4(12):2147–2157. [PubMed]
  • Brewin NJ. Development of the legume root nodule. Annu Rev Cell Biol. 1991;7:191–226. [PubMed]
  • Charles TC, Newcomb W, Finan TM. ndvF, a novel locus located on megaplasmid pRmeSU47b (pEXO) of Rhizobium meliloti, is required for normal nodule development. J Bacteriol. 1991 Jul;173(13):3981–3992. [PMC free article] [PubMed]
  • De Vos GF, Walker GC, Signer ER. Genetic manipulations in Rhizobium meliloti utilizing two new transposon Tn5 derivatives. Mol Gen Genet. 1986 Sep;204(3):485–491. [PubMed]
  • Dickstein R, Scheirer DC, Fowle WH, Ausubel FM. Nodules elicited by Rhizobium meliloti heme mutants are arrested at an early stage of development. Mol Gen Genet. 1991 Dec;230(3):423–432. [PubMed]
  • Ditta G, Schmidhauser T, Yakobson E, Lu P, Liang XW, Finlay DR, Guiney D, Helinski DR. Plasmids related to the broad host range vector, pRK290, useful for gene cloning and for monitoring gene expression. Plasmid. 1985 Mar;13(2):149–153. [PubMed]
  • Dylan T, Ielpi L, Stanfield S, Kashyap L, Douglas C, Yanofsky M, Nester E, Helinski DR, Ditta G. Rhizobium meliloti genes required for nodule development are related to chromosomal virulence genes in Agrobacterium tumefaciens. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4403–4407. [PMC free article] [PubMed]
  • Elzer PH, Phillips RW, Kovach ME, Peterson KM, Roop RM., 2nd Characterization and genetic complementation of a Brucella abortus high-temperature-requirement A (htrA) deletion mutant. Infect Immun. 1994 Oct;62(10):4135–4139. [PMC free article] [PubMed]
  • Finan TM, Hartweig E, LeMieux K, Bergman K, Walker GC, Signer ER. General transduction in Rhizobium meliloti. J Bacteriol. 1984 Jul;159(1):120–124. [PMC free article] [PubMed]
  • Finan TM, Hirsch AM, Leigh JA, Johansen E, Kuldau GA, Deegan S, Walker GC, Signer ER. Symbiotic mutants of Rhizobium meliloti that uncouple plant from bacterial differentiation. Cell. 1985 Apr;40(4):869–877. [PubMed]
  • Fisher RF, Long SR. Rhizobium--plant signal exchange. Nature. 1992 Jun 25;357(6380):655–660. [PubMed]
  • Friedman AM, Long SR, Brown SE, Buikema WJ, Ausubel FM. Construction of a broad host range cosmid cloning vector and its use in the genetic analysis of Rhizobium mutants. Gene. 1982 Jun;18(3):289–296. [PubMed]
  • Geremia RA, Cavaignac S, Zorreguieta A, Toro N, Olivares J, Ugalde RA. A Rhizobium meliloti mutant that forms ineffective pseudonodules in alfalfa produces exopolysaccharide but fails to form beta-(1----2) glucan. J Bacteriol. 1987 Feb;169(2):880–884. [PMC free article] [PubMed]
  • Glazebrook J, Ichige A, Walker GC. A Rhizobium meliloti homolog of the Escherichia coli peptide-antibiotic transport protein SbmA is essential for bacteroid development. Genes Dev. 1993 Aug;7(8):1485–1497. [PubMed]
  • Glazebrook J, Meiri G, Walker GC. Genetic mapping of symbiotic loci on the Rhizobium meliloti chromosome. Mol Plant Microbe Interact. 1992 May-Jun;5(3):223–227. [PubMed]
  • Glazebrook J, Walker GC. A novel exopolysaccharide can function in place of the calcofluor-binding exopolysaccharide in nodulation of alfalfa by Rhizobium meliloti. Cell. 1989 Feb 24;56(4):661–672. [PubMed]
  • Glucksmann MA, Reuber TL, Walker GC. Family of glycosyl transferases needed for the synthesis of succinoglycan by Rhizobium meliloti. J Bacteriol. 1993 Nov;175(21):7033–7044. [PMC free article] [PubMed]
  • Glucksmann MA, Reuber TL, Walker GC. Genes needed for the modification, polymerization, export, and processing of succinoglycan by Rhizobium meliloti: a model for succinoglycan biosynthesis. J Bacteriol. 1993 Nov;175(21):7045–7055. [PMC free article] [PubMed]
  • Hirsch AM, Bang M, Ausubel FM. Ultrastructural analysis of ineffective alfalfa nodules formed by nif::Tn5 mutants of Rhizobium meliloti. J Bacteriol. 1983 Jul;155(1):367–380. [PMC free article] [PubMed]
  • Hirsch AM, Smith CA. Effects of Rhizobium meliloti nif and fix mutants on alfalfa root nodule development. J Bacteriol. 1987 Mar;169(3):1137–1146. [PMC free article] [PubMed]
  • Johnson K, Charles I, Dougan G, Pickard D, O'Gaora P, Costa G, Ali T, Miller I, Hormaeche C. The role of a stress-response protein in Salmonella typhimurium virulence. Mol Microbiol. 1991 Feb;5(2):401–407. [PubMed]
  • Jurtshuk P, Jr, Mueller TJ, Acord WC. Bacterial terminal oxidases. CRC Crit Rev Microbiol. 1975 May;3(4):399–468. [PubMed]
  • Kereszt A, Slaska-Kiss K, Putnoky P, Banfalvi Z, Kondorosi A. The cycHJKL genes of Rhizobium meliloti involved in cytochrome c biogenesis are required for "respiratory" nitrate reduction ex planta and for nitrogen fixation during symbiosis. Mol Gen Genet. 1995 Apr 10;247(1):39–47. [PubMed]
  • Leigh JA, Reed JW, Hanks JF, Hirsch AM, Walker GC. Rhizobium meliloti mutants that fail to succinylate their calcofluor-binding exopolysaccharide are defective in nodule invasion. Cell. 1987 Nov 20;51(4):579–587. [PubMed]
  • Leigh JA, Signer ER, Walker GC. Exopolysaccharide-deficient mutants of Rhizobium meliloti that form ineffective nodules. Proc Natl Acad Sci U S A. 1985 Sep;82(18):6231–6235. [PMC free article] [PubMed]
  • Lipinska B, Fayet O, Baird L, Georgopoulos C. Identification, characterization, and mapping of the Escherichia coli htrA gene, whose product is essential for bacterial growth only at elevated temperatures. J Bacteriol. 1989 Mar;171(3):1574–1584. [PMC free article] [PubMed]
  • Lipinska B, Sharma S, Georgopoulos C. Sequence analysis and regulation of the htrA gene of Escherichia coli: a sigma 32-independent mechanism of heat-inducible transcription. Nucleic Acids Res. 1988 Nov 11;16(21):10053–10067. [PMC free article] [PubMed]
  • Lipinska B, Zylicz M, Georgopoulos C. The HtrA (DegP) protein, essential for Escherichia coli survival at high temperatures, is an endopeptidase. J Bacteriol. 1990 Apr;172(4):1791–1797. [PMC free article] [PubMed]
  • Liu J. Microcin B17: posttranslational modifications and their biological implications. Proc Natl Acad Sci U S A. 1994 May 24;91(11):4618–4620. [PMC free article] [PubMed]
  • Loferer H, Bott M, Hennecke H. Bradyrhizobium japonicum TlpA, a novel membrane-anchored thioredoxin-like protein involved in the biogenesis of cytochrome aa3 and development of symbiosis. EMBO J. 1993 Sep;12(9):3373–3383. [PMC free article] [PubMed]
  • Long S, McCune S, Walker GC. Symbiotic loci of Rhizobium meliloti identified by random TnphoA mutagenesis. J Bacteriol. 1988 Sep;170(9):4257–4265. [PMC free article] [PubMed]
  • Long S, Reed JW, Himawan J, Walker GC. Genetic analysis of a cluster of genes required for synthesis of the calcofluor-binding exopolysaccharide of Rhizobium meliloti. J Bacteriol. 1988 Sep;170(9):4239–4248. [PMC free article] [PubMed]
  • Manoil C, Beckwith J. TnphoA: a transposon probe for protein export signals. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129–8133. [PMC free article] [PubMed]
  • Meade HM, Long SR, Ruvkun GB, Brown SE, Ausubel FM. Physical and genetic characterization of symbiotic and auxotrophic mutants of Rhizobium meliloti induced by transposon Tn5 mutagenesis. J Bacteriol. 1982 Jan;149(1):114–122. [PMC free article] [PubMed]
  • Preisig O, Anthamatten D, Hennecke H. Genes for a microaerobically induced oxidase complex in Bradyrhizobium japonicum are essential for a nitrogen-fixing endosymbiosis. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3309–3313. [PMC free article] [PubMed]
  • Ramseier TM, Winteler HV, Hennecke H. Discovery and sequence analysis of bacterial genes involved in the biogenesis of c-type cytochromes. J Biol Chem. 1991 Apr 25;266(12):7793–7803. [PubMed]
  • Ritz D, Bott M, Hennecke H. Formation of several bacterial c-type cytochromes requires a novel membrane-anchored protein that faces the periplasm. Mol Microbiol. 1993 Aug;9(4):729–740. [PubMed]
  • Ritz D, Thöny-Meyer L, Hennecke H. The cycHJKL gene cluster plays an essential role in the biogenesis of c-type cytochromes in Bradyrhizobium japonicum. Mol Gen Genet. 1995 Apr 10;247(1):27–38. [PubMed]
  • Roop RM, 2nd, Fletcher TW, Sriranganathan NM, Boyle SM, Schurig GG. Identification of an immunoreactive Brucella abortus HtrA stress response protein homolog. Infect Immun. 1994 Mar;62(3):1000–1007. [PMC free article] [PubMed]
  • Stachel SE, An G, Flores C, Nester EW. A Tn3 lacZ transposon for the random generation of beta-galactosidase gene fusions: application to the analysis of gene expression in Agrobacterium. EMBO J. 1985 Apr;4(4):891–898. [PMC free article] [PubMed]
  • Strauch KL, Beckwith J. An Escherichia coli mutation preventing degradation of abnormal periplasmic proteins. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1576–1580. [PMC free article] [PubMed]
  • Strauch KL, Johnson K, Beckwith J. Characterization of degP, a gene required for proteolysis in the cell envelope and essential for growth of Escherichia coli at high temperature. J Bacteriol. 1989 May;171(5):2689–2696. [PMC free article] [PubMed]
  • Thöny-Meyer L, Ritz D, Hennecke H. Cytochrome c biogenesis in bacteria: a possible pathway begins to emerge. Mol Microbiol. 1994 Apr;12(1):1–9. [PubMed]
  • Thöny-Meyer L, Stax D, Hennecke H. An unusual gene cluster for the cytochrome bc1 complex in Bradyrhizobium japonicum and its requirement for effective root nodule symbiosis. Cell. 1989 May 19;57(4):683–697. [PubMed]
  • Yarosh OK, Charles TC, Finan TM. Analysis of C4-dicarboxylate transport genes in Rhizobium meliloti. Mol Microbiol. 1989 Jun;3(6):813–823. [PubMed]
  • Yorgey P, Lee J, Kördel J, Vivas E, Warner P, Jebaratnam D, Kolter R. Posttranslational modifications in microcin B17 define an additional class of DNA gyrase inhibitor. Proc Natl Acad Sci U S A. 1994 May 10;91(10):4519–4523. [PMC free article] [PubMed]

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

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...