Activity of the Agrobacterium Ti plasmid conjugal transfer regulator TraR is inhibited by the product of the traM gene
10.1128/jb.177.5.1367-1373.1995
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

The Agrobacterium Ti plasmid tra regulon was previously found to be positively regulated by the TraR protein in the presence of a diffusible N-acyl homoserine lactone designated Agrobacterium autoinducer (AAI). TraR and AAI are similar to LuxR from Vibrio fischeri and the Vibrio autoinducer (VAI), which regulate target bioluminescence (lux) genes in a cell density-dependent manner. We now show that tra genes are also regulated by a second protein, designated TraM, which acts to antagonize TraR-dependent activation. The traM gene is closely linked to traR, and the two genes are transcribed convergently. The predicted TraM proteins of two different Ti plasmids are 77% identical but are not significantly similar to other protein sequences in the database, and thus TraM may represent a novel regulatory protein. Null mutations in traM cause strongly increased conjugation, tra gene transcription, and AAI production. A functional copy of traM introduced into traM mutants decreased conjugation, tra gene transcription, and AAI synthesis. TraM inhibits transcription of traA, traI, and traM. Although traM was first identified by its octopine-inducible promoter, we now show that induction by octopine requires traR, strongly suggesting that TraR is the direct traM activator.

Bibliography

Fuqua, C., Burbea, M., & Winans, S. C. (1995). Activity of the Agrobacterium Ti plasmid conjugal transfer regulator TraR is inhibited by the product of the traM gene. Journal of Bacteriology, 177(5), 1367–1373.

Authors 3
  1. C Fuqua (first)
  2. M Burbea (additional)
  3. S C Winans (additional)
References 49 Referenced 123
  1. Altschul , S. F. , W. Gish , W. Miller , E. W. Myers , and D. J. Lipman . 1990 . Basic local alignment search tool . J. Mol. Biol. 215 : 403 - 410 . (10.1016/S0022-2836(05)80360-2) / J. Mol. Biol. / Basic local alignment search tool by Altschul S. F. (1990)
  2. Bassler , B. L. , M. Wright , and M. R. Silverman . 1994 . Multiple signalling systems controlling expression of Iuminescence in Vibrio harveyi: sequence and function of genes encoding a second sensory pathway . Mol. Microbiol. 13 : 273 - 286 . (10.1111/j.1365-2958.1994.tb00422.x) / Mol. Microbiol. / Multiple signalling systems controlling expression of Iuminescence in Vibrio harveyi: sequence and function of genes encoding a second sensory pathway by Bassler B. L. (1994)
  3. Bassler , B. L. , M. Wright , and M. R. Silverman . 1994 . Sequence and function of LuxO, a negative regulator of Iuminescence in Vibrio harveyi . Mol. Microbiol. 12 : 403 - 412 . (10.1111/j.1365-2958.1994.tb01029.x) / Mol. Microbiol. / Sequence and function of LuxO, a negative regulator of Iuminescence in Vibrio harveyi by Bassler B. L. (1994)
  4. Beck von Bodman , S. , G. T. Hayman , and S. K. Farrand . 1992 . Opine catabolism and conjugal transfer of the nopaline Ti plasmid pTiC58 are coordinately regulated by a single repressor . Proc. Natl. Acad. Sci. USA 89 : 643 - 647 . (10.1073/pnas.89.2.643) / Proc. Natl. Acad. Sci. USA / Opine catabolism and conjugal transfer of the nopaline Ti plasmid pTiC58 are coordinately regulated by a single repressor by Beck von Bodman S. (1992)
  5. Bukau , B. 1993 . Regulation of the Escherichia coli heat-shock response . Mol. Microbiol. 9 : 671 - 680 . (10.1111/j.1365-2958.1993.tb01727.x) / Mol. Microbiol. / Regulation of the Escherichia coli heat-shock response by Bukau B. (1993)
  6. Chen , C. - Y. , and S. C. Winans . 1991 . Controlled expression of the transcriptional activator gene virG in Agrobacterium tumefaciens by using the Escherichia coli lac promoter . J. Bacteriol. 173 : 1139 - 1144 . (10.1128/JB.173.3.1139-1144.1991) / J. Bacteriol. / Controlled expression of the transcriptional activator gene virG in Agrobacterium tumefaciens by using the Escherichia coli lac promoter by Chen C. (1991)
  7. Cho , K. - Y. , and S. C. Winans . 1993 . Altered-function mutations in the Agrobacterium tumefaciens OccR protein and in an OccR-regulated promoter . J. Bacteriol. 175 : 7715 - 7719 . (10.1128/JB.175.23.7715-7719.1993) / J. Bacteriol. / Altered-function mutations in the Agrobacterium tumefaciens OccR protein and in an OccR-regulated promoter by Cho K. (1993)
  8. Cho K.-Y. and S. C. Winans. Unpublished results.
  9. Cubo , M. T. , A. Economou , G. Murphy , A. W. B. Johnston , and J. A. Downie . 1992 . Molecular characterization and regulation of the rhizosphere-expressed genes rhiABCR that can influence nodulation by Rhizobium leguminosarum biovar viciae . J. Bacteriol. 174 : 4026 - 4035 . (10.1128/JB.174.12.4026-4035.1992) / J. Bacteriol. / Molecular characterization and regulation of the rhizosphere-expressed genes rhiABCR that can influence nodulation by Rhizobium leguminosarum biovar viciae by Cubo M. T. (1992)
  10. Devereux , J. , P. Haeberli , and O. Smithies . 1984 . A comprehensive set of sequence analysis programs for the VAX . Nucleic Acids Res. 12 : 387 - 395 . (10.1093/nar/12.1Part1.387) / Nucleic Acids Res. / A comprehensive set of sequence analysis programs for the VAX by Devereux J. (1984)
  11. De Vos , G. , M. De Beuckeleer , M. Von Montagu , and J. Schell . 1981 . Restriction endonuclease mapping of the octopine tumor-inducing plasmid pTiAch5 of Agrobacterium tumefaciens . Plasmid 6 : 249 - 253 . (10.1016/0147-619X(81)90070-6) / Plasmid / Restriction endonuclease mapping of the octopine tumor-inducing plasmid pTiAch5 of Agrobacterium tumefaciens by De Vos G. (1981)
  12. Dunlap , P. V. 1989 . Regulation of Iuminescence by cyclic AMP in cya-like and crp-like mutants of Vibrio fischeri . J. Bacteriol. 171 : 1199 - 1202 . (10.1128/jb.171.2.1199-1202.1989) / J. Bacteriol. / Regulation of Iuminescence by cyclic AMP in cya-like and crp-like mutants of Vibrio fischeri by Dunlap P. V. (1989)
  13. Dunlap , P. V. , and E. P. Greenberg . 1985 . Control of Vibrio fischeri Iuminescence genes in Escherichia coli by cyclic AMP and cyclic AMP receptor protein . J. Bacteriol. 164 : 45 - 50 . (10.1128/JB.164.1.45-50.1985) / J. Bacteriol. / Control of Vibrio fischeri Iuminescence genes in Escherichia coli by cyclic AMP and cyclic AMP receptor protein by Dunlap P. V. (1985)
  14. Dunlap , P. V. , and E. P. Greenberg . 1988 . Analysis of the mechanism of Vibrio fischeri Iuminescence gene regulation by cyclic AMP and cyclic AMP receptor protein in Escherichia coli . J. Bacteriol. 170 : 4040 - 4046 . (10.1128/JB.170.9.4040-4046.1988) / J. Bacteriol. / Analysis of the mechanism of Vibrio fischeri Iuminescence gene regulation by cyclic AMP and cyclic AMP receptor protein in Escherichia coli by Dunlap P. V. (1988)
  15. Ellis , J. G. , P. J. Murphy , and A. Kerr . 1982 . Isolation and properties of transfer regulatory mutants of the nopaline Ti plasmid pTiC58 . Mol. Gen. Genet. 186 : 275 - 281 . (10.1007/BF00331862) / Mol. Gen. Genet. / Isolation and properties of transfer regulatory mutants of the nopaline Ti plasmid pTiC58 by Ellis J. G. (1982)
  16. Engler , G. , A. Depicker , R. Maenhaut , R. Villarroel , M. Van Montagu , and J. Schell . 1981 . Physical mapping of DNA base sequence homologies between an octopine and a nopaline Ti plasmid of Agrobacterium tumefaciens . J. Mol. Biol. 152 : 183 - 208 . (10.1016/0022-2836(81)90239-4) / J. Mol. Biol. / Physical mapping of DNA base sequence homologies between an octopine and a nopaline Ti plasmid of Agrobacterium tumefaciens by Engler G. (1981)
  17. Farrand S. K. 1993. Conjugal transfer of Agrobacterium plasmids p. 255-291. In D. B. Clewell (ed.) Bacterial conjugation. Plenum Publishing Corp. New York. (10.1007/978-1-4757-9357-4_10)
  18. Farrand S. K. Personal communications.
  19. Fuqua , W. C. , and S. C. Winans . 1994 . A LuxR-LuxI type regulatory system activates Agrobacterium Ti plasmid conjugal transfer in the presence of a plant tumor metabolite . J. Bacteriol. 176 : 2796 - 2806 . (10.1128/JB.176.10.2796-2806.1994) / J. Bacteriol. / A LuxR-LuxI type regulatory system activates Agrobacterium Ti plasmid conjugal transfer in the presence of a plant tumor metabolite by Fuqua W. C. (1994)
  20. Fuqua W. C. and S. C. Winans. Unpublished results.
  21. Fuqua , W. C. , S. C. Winans , and E. P. Greenberg . 1994 . Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators . J. Bacteriol. 176 : 269 - 275 . (10.1128/JB.176.2.269-275.1994) / J. Bacteriol. / Quorum sensing in bacteria: the LuxR-LuxI family of cell density-responsive transcriptional regulators by Fuqua W. C. (1994)
  22. Gelfand D. H. J. J. Sninsky and T. J. White. 1990. Optimization of PCR p. 3-12. In M. A. Innis and D. H. Gelfand (ed.) PCR protocols: a guide to methods and applications. Academic Press New York. (10.1016/B978-0-12-372180-8.50005-6)
  23. Genetello , C. , N. Van Larebeke , M. Holsters , A. De Picker , M. Van Montagu , and J. Schell . 1977 . Ti plasmids of Agrobacterium as conjugative plasmids . Nature (London) 265 : 561 - 563 . (10.1038/265561a0) / Nature (London) / Ti plasmids of Agrobacterium as conjugative plasmids by Genetello C. (1977)
  24. Gray , K. M. , L. Passador , B. H. Iglewski , and E. P. Greenberg . 1994 . Interchangeability and specificity of components from the quorum-sensing regulatory systems of Vibrio fischeri and Pseudomonas aeruginosa . J. Bacteriol. 176 : 3076 - 3080 . (10.1128/JB.176.10.3076-3080.1994) / J. Bacteriol. / Interchangeability and specificity of components from the quorum-sensing regulatory systems of Vibrio fischeri and Pseudomonas aeruginosa by Gray K. M. (1994)
  25. Habeeb , L. F. , L. Wang , and S. C. Winans . 1991 . Transcription of the octopine catabolism operon of the Agrobacterium tumor-inducing plasmid pTiA6 is activated by a LysR homolog . Mol. Plant Microbe Interact. 4 : 379 - 385 . (10.1094/MPMI-4-379) / Mol. Plant Microbe Interact. / Transcription of the octopine catabolism operon of the Agrobacterium tumor-inducing plasmid pTiA6 is activated by a LysR homolog by Habeeb L. F. (1991)
  26. Han , D. C. , C. -Y. Chen , Y. -F. Chen , and S. C. Winans . 1992 . Alteredfunction mutations of the transcriptional regulatory gene virG of Agrobacterium tumefaciens . J. Bacteriol. 174 : 7040 - 7043 . (10.1128/JB.174.21.7040-7043.1992) / J. Bacteriol. / Alteredfunction mutations of the transcriptional regulatory gene virG of Agrobacterium tumefaciens by Han D. C. (1992)
  27. Hwang , I. , D. M. Cook , and S. K. Farrand . 1995 . A new regulatory element modulates homoserine lactone-mediated autoinduction of Ti plasmid conjugal transfer . J. Bacteriol. 177 : 449 - 458 . (10.1128/jb.177.2.449-458.1995) / J. Bacteriol. / A new regulatory element modulates homoserine lactone-mediated autoinduction of Ti plasmid conjugal transfer by Hwang I. (1995)
  28. Hwang , I. , P. -L. Li , L. Zhang , K. R. Piper , D. M. Cook , M. E. Tate , and S. K. Farrand . 1994 . TraI, a LuxI homologue, is responsible for production of conjugation factor, the Ti plasmid N-acylhomoserine lactone autoinducer . Proc. Natl. Acad. Sci. USA 91 : 4639 - 4643 . (10.1073/pnas.91.11.4639) / Proc. Natl. Acad. Sci. USA / TraI, a LuxI homologue, is responsible for production of conjugation factor, the Ti plasmid N-acylhomoserine lactone autoinducer by Hwang I. (1994)
  29. Kaniga , K. , I. Delor , and G. R. Cornelis . 1991 . A wide-host-range suicide vector for improving reverse genetics in Gram-negative bacteria: inactivation of the blaA gene of Yersinia enterocolitica . Gene 109 : 137 - 141 . (10.1016/0378-1119(91)90599-7) / Gene / A wide-host-range suicide vector for improving reverse genetics in Gram-negative bacteria: inactivation of the blaA gene of Yersinia enterocolitica by Kaniga K. (1991)
  30. Kaplan , H. B. , and E. P. Greenberg . 1985 . Diffusion of autoinducer is involved in regulation of the Vibrio fischeri Iuminescence system . J. Bacteriol. 163 : 1210 - 1214 . (10.1128/JB.163.3.1210-1214.1985) / J. Bacteriol. / Diffusion of autoinducer is involved in regulation of the Vibrio fischeri Iuminescence system by Kaplan H. B. (1985)
  31. Kerr , A. , P. Manigault , and J. Tempé . 1977 . Transfer of virulence in vivo and in vitro in Agrobacterium . Nature (London) 265 : 560 - 561 . (10.1038/265560a0) / Nature (London) / Transfer of virulence in vivo and in vitro in Agrobacterium by Kerr A. (1977)
  32. Klapwijk , P. M. , T. Scheulderman , and R. Schilperoort . 1978 . Coordinated regulation of octopine degradation and conjugative transfer of Ti plasmids in Agrobacterium tumefaciens: evidence for a common regulatory gene and separate operons . J. Bacteriol. 136 : 775 - 785 . (10.1128/JB.136.2.775-785.1978) / J. Bacteriol. / Coordinated regulation of octopine degradation and conjugative transfer of Ti plasmids in Agrobacterium tumefaciens: evidence for a common regulatory gene and separate operons by Klapwijk P. M. (1978)
  33. McBride , K. E. , and K. R. Summerfelt . 1990 . Improved binary vectors for Agrobacterium-mediated plant transformation . Plant Mol. Biol. 14 : 269 - 276 . (10.1007/BF00018567) / Plant Mol. Biol. / Improved binary vectors for Agrobacterium-mediated plant transformation by McBride K. E. (1990)
  34. Mersereau , M. , G. J. Pazour , and A. Das . 1990 . Efficient transformation of Agrobacterium tumefaciens by electroporation . Gene 90 : 149 - 151 . (10.1016/0378-1119(90)90452-W) / Gene / Efficient transformation of Agrobacterium tumefaciens by electroporation by Mersereau M. (1990)
  35. Messing , J. 1979 . A multipurpose cloning system based on single-stranded bacteriophage M13 . Recomb. DNA Tech. Bull. 2 : 43 - 48 . / Recomb. DNA Tech. Bull. / A multipurpose cloning system based on single-stranded bacteriophage M13 by Messing J. (1979)
  36. Miller , V. L. , and J. J. Mekalanos . 1988 . A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae . J. Bacteriol. 170 : 2575 - 2583 . (10.1128/JB.170.6.2575-2583.1988) / J. Bacteriol. / A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae by Miller V. L. (1988)
  37. Moré M. I. and S. C. Winans. Unpublished results.
  38. Pearson , W. R. , and D. J. Lipman . 1988 . Improved tools for biological sequence comparison . Proc. Natl. Acad. Sci. USA 85 : 2444 - 2448 . (10.1073/pnas.85.8.2444) / Proc. Natl. Acad. Sci. USA / Improved tools for biological sequence comparison by Pearson W. R. (1988)
  39. Piper , K. R. , S. Beck von Bodman , and S. K. Farrand . 1993 . Conjugation factor of Agrobacterium tumefaciens regulates Ti plasmid transfer by autoinduction . Nature (London) 362 : 448 - 450 . (10.1038/362448a0) / Nature (London) / Conjugation factor of Agrobacterium tumefaciens regulates Ti plasmid transfer by autoinduction by Piper K. R. (1993)
  40. Rosson , R. A. , and K. H. Nealson . 1981 . Autoinduction of bacterial bioluminescence in a carbon limited chemostat . Arch. Microbiol. 129 : 299 - 304 . (10.1007/BF00414701) / Arch. Microbiol. / Autoinduction of bacterial bioluminescence in a carbon limited chemostat by Rosson R. A. (1981)
  41. Sambrook J. E. F. Fritsch and T. Maniatis. 1989. Molecular cloning: a laboratory manual 2nd ed. Cold Spring Harbor Laboratory Press Cold Spring Harbor N.Y.
  42. Sciaky , D. , A. L. Montoya , and M. -D. Chilton . 1977 . Fingerprints of Agrobacterium Ti plasmids . Plasmid 1 : 238 - 253 . (10.1016/0147-619X(78)90042-2) / Plasmid / Fingerprints of Agrobacterium Ti plasmids by Sciaky D. (1977)
  43. Sharma , S. B. , and E. R. Signer . 1990 . Temporal and spatial regulation of the symbiotic genes of Rhizobium meliloti in planta revealed by transposon Tn5-gusA . Genes Dev. 4 : 344 - 356 . (10.1101/gad.4.3.344) / Genes Dev. / Temporal and spatial regulation of the symbiotic genes of Rhizobium meliloti in planta revealed by transposon Tn5-gusA by Sharma S. B. (1990)
  44. Swift , S. , M. K. Winson , P. F. Chan , N. J. Bainton , M. Birdsall , P. J. Reeves , C. E. D. Rees , S. R. Chhabra , P. J. Hill , J. P. Throup , B. W. Bycroft , G. P. C. Salmond , P. Williams , and G. S. A. B. Stewart . 1993 . A novel strategy for the isolation of luxI homologues: evidence for the widespread distribution of a LuxR:LuxI superfamily in enteric bacteria . Mol. Microbiol. 10 : 511 - 520 . (10.1111/j.1365-2958.1993.tb00923.x) / Mol. Microbiol. / A novel strategy for the isolation of luxI homologues: evidence for the widespread distribution of a LuxR:LuxI superfamily in enteric bacteria by Swift S. (1993)
  45. Tempé , J. , A. Petit , M. Holsters , M. Van Montagu , and J. Schell . 1977 . Thermosensitive step associated with transfer of the Ti plasmid during conjugation: possible relation to transformation in crown gall . Proc. Natl. Acad. Sci. USA 74 : 2848 - 2849 . (10.1073/pnas.74.7.2848) / Proc. Natl. Acad. Sci. USA / Thermosensitive step associated with transfer of the Ti plasmid during conjugation: possible relation to transformation in crown gall by Tempé J. (1977)
  46. Walker G. C. 1987. The SOS response of Escherichia coli p. 1346-1357. In F. C. Neidhardt J. L. Ingraham K. B. Low B. Magasanik M. Schaechter and H. E. Umbarger (ed.) Escherichia coli and Salmonella typhimurium: cellular and molecular biology. American Society for Microbiology Washington D.C.
  47. Wang , L. , J. D. Helmann , and S. C. Winans . 1992 . The Agrobacterium tumefaciens transcriptional activator OccR causes a bend at a target promoter, which is partially relaxed by a plant tumor metabolite . Cell 69 : 659 - 667 . (10.1016/0092-8674(92)90229-6) / Cell / The Agrobacterium tumefaciens transcriptional activator OccR causes a bend at a target promoter, which is partially relaxed by a plant tumor metabolite by Wang L. (1992)
  48. Winans , S. C. , N. J. Mantis , C. -Y. Chen , C. -H. Chang , and D. C. Han . 1994 . Host recognition by the VirA, VirG two-component regulatory proteins of Agrobacterium tumefaciens . Res. Microbiol. 145 : 345 - 496 . (10.1016/0923-2508(94)90095-7) / Res. Microbiol. / Host recognition by the VirA, VirG two-component regulatory proteins of Agrobacterium tumefaciens by Winans S. C. (1994)
  49. Zhang , L. , P. J. Murphy , A. Kerr , and M. E. Tate . 1993 . Agrobacterium conjugation and gene regulation by N-acyl-homoserine lactones . Nature (London) 362 : 446 - 448 . (10.1038/362446a0) / Nature (London) / Agrobacterium conjugation and gene regulation by N-acyl-homoserine lactones by Zhang L. (1993)
Dates
Type When
Created 8 years, 9 months ago (Nov. 9, 2016, 11:10 a.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 1:54 p.m.)
Indexed 3 weeks, 3 days ago (Aug. 5, 2025, 8:52 a.m.)
Issued 30 years, 5 months ago (March 1, 1995)
Published 30 years, 5 months ago (March 1, 1995)
Published Print 30 years, 5 months ago (March 1, 1995)
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@article{Fuqua_1995, title={Activity of the Agrobacterium Ti plasmid conjugal transfer regulator TraR is inhibited by the product of the traM gene}, volume={177}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.177.5.1367-1373.1995}, DOI={10.1128/jb.177.5.1367-1373.1995}, number={5}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Fuqua, C and Burbea, M and Winans, S C}, year={1995}, month=mar, pages={1367–1373} }