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Membrane topology and functional analysis of the sensory protein VirA of Agrobacterium tumefaciens.

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  • 1. Department of Plant Molecular Biology, Leiden University, The Netherlands.
      Authors
    • Melchers LS 1
    (1 author)

ORCIDs linked to this article

The EMBO Journal, 01 Jul 1989, 8(7):1919-1925
https://doi.org/10.1002/j.1460-2075.1989.tb03595.x PMID: 2792074 PMCID: PMC401051

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Abstract 

The virA gene of Agrobacterium tumefaciens encodes an inner membrane that mediates the transcriptional activation of virulence genes in response to plant signal molecules. We report here a functional analysis of the N-terminal, C-terminal and periplasmic domains of VirA in transmembrane signalling. First, we show that VirA has a transmembrane topology by analysis of the alkaline phosphatase activities, determined by several virA-phoA gene fusions. Second, we report here the construction of several virA-tar chimeric genes, in which the 3'-coding region of virA is conserved to study transmembrane signalling, as well as the construction of a set of virA deletion mutations. Results of analyses of vir induction behaviour and tumour inducing abilities of agrobacteria carrying these mutant genes do not support existing models for the chemoreceptor function of the VirA periplasmic domain. We demonstrate that the periplasmic domain of VirA can be either replaced by a corresponding region of the E.coli chemosensory protein Tar or even totally deleted from VirA without a loss of function. Here, we present a model of VirA which involves a receptor function for the second membrane-spanning domain and an intracellular signalling function for the cytoplasmic domain of VirA. In addition, we show that VirA plays a role in determining the sensitivity for pH and temperature in acetosyringone-mediated vir induction, and we propose a role for the VirA periplasmic domain in detection of the external pH conditions.

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EMBO J. 1989 Jul; 8(7): 1919–1925.
PMCID: PMC401051
PMID: 2792074

Membrane topology and functional analysis of the sensory protein VirA of Agrobacterium tumefaciens.

L S Melchers, T J Regensburg-Tuïnk, R B Bourret, N J Sedee, R A Schilperoort, and P J Hooykaas
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Abstract

The virA gene of Agrobacterium tumefaciens encodes an inner membrane that mediates the transcriptional activation of virulence genes in response to plant signal molecules. We report here a functional analysis of the N-terminal, C-terminal and periplasmic domains of VirA in transmembrane signalling. First, we show that VirA has a transmembrane topology by analysis of the alkaline phosphatase activities, determined by several virA-phoA gene fusions. Second, we report here the construction of several virA-tar chimeric genes, in which the 3'-coding region of virA is conserved to study transmembrane signalling, as well as the construction of a set of virA deletion mutations. Results of analyses of vir induction behaviour and tumour inducing abilities of agrobacteria carrying these mutant genes do not support existing models for the chemoreceptor function of the VirA periplasmic domain. We demonstrate that the periplasmic domain of VirA can be either replaced by a corresponding region of the E.coli chemosensory protein Tar or even totally deleted from VirA without a loss of function. Here, we present a model of VirA which involves a receptor function for the second membrane-spanning domain and an intracellular signalling function for the cytoplasmic domain of VirA. In addition, we show that VirA plays a role in determining the sensitivity for pH and temperature in acetosyringone-mediated vir induction, and we propose a role for the VirA periplasmic domain in detection of the external pH conditions.

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Selected References

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  • Adams GA, Rose JK. Structural requirements of a membrane-spanning domain for protein anchoring and cell surface transport. Cell. 1985 Jul;41(3):1007–1015. [Abstract] [Google Scholar]
  • Ames P, Parkinson JS. Transmembrane signaling by bacterial chemoreceptors: E. coli transducers with locked signal output. Cell. 1988 Dec 2;55(5):817–826. [Abstract] [Google Scholar]
  • Carter P, Bedouelle H, Winter G. Improved oligonucleotide site-directed mutagenesis using M13 vectors. Nucleic Acids Res. 1985 Jun 25;13(12):4431–4443. [Europe PMC free article] [Abstract] [Google Scholar]
  • Chen EY, Seeburg PH. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. [Abstract] [Google Scholar]
  • Figurski DH, Helinski DR. Replication of an origin-containing derivative of plasmid RK2 dependent on a plasmid function provided in trans. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1648–1652. [Europe PMC free article] [Abstract] [Google Scholar]
  • Forst S, Comeau D, Norioka S, Inouye M. Localization and membrane topology of EnvZ, a protein involved in osmoregulation of OmpF and OmpC in Escherichia coli. J Biol Chem. 1987 Dec 5;262(34):16433–16438. [Abstract] [Google Scholar]
  • Gebert JF, Overhoff B, Manson MD, Boos W. The Tsr chemosensory transducer of Escherichia coli assembles into the cytoplasmic membrane via a SecA-dependent process. J Biol Chem. 1988 Nov 15;263(32):16652–16660. [Abstract] [Google Scholar]
  • Giphart-Gassler M, Van de Putte P. Thermo-inducible expression of cloned early genes of bacteriophage Mu. Gene. 1979 Sep;7(1):33–50. [Abstract] [Google Scholar]
  • Hess JF, Oosawa K, Matsumura P, Simon MI. Protein phosphorylation is involved in bacterial chemotaxis. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7609–7613. [Europe PMC free article] [Abstract] [Google Scholar]
  • Hoffman CS, Wright A. Fusions of secreted proteins to alkaline phosphatase: an approach for studying protein secretion. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5107–5111. [Europe PMC free article] [Abstract] [Google Scholar]
  • Igo MM, Silhavy TJ. EnvZ, a transmembrane environmental sensor of Escherichia coli K-12, is phosphorylated in vitro. J Bacteriol. 1988 Dec;170(12):5971–5973. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kofoid EC, Parkinson JS. Transmitter and receiver modules in bacterial signaling proteins. Proc Natl Acad Sci U S A. 1988 Jul;85(14):4981–4985. [Europe PMC free article] [Abstract] [Google Scholar]
  • Krikos A, Mutoh N, Boyd A, Simon MI. Sensory transducers of E. coli are composed of discrete structural and functional domains. Cell. 1983 Jun;33(2):615–622. [Abstract] [Google Scholar]
  • Leroux B, Yanofsky MF, Winans SC, Ward JE, Ziegler SF, Nester EW. Characterization of the virA locus of Agrobacterium tumefaciens: a transcriptional regulator and host range determinant. EMBO J. 1987 Apr;6(4):849–856. [Europe PMC free article] [Abstract] [Google Scholar]
  • 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Manoil C, Beckwith J. A genetic approach to analyzing membrane protein topology. Science. 1986 Sep 26;233(4771):1403–1408. [Abstract] [Google Scholar]
  • Melchers LS, Thompson DV, Idler KB, Schilperoort RA, Hooykaas PJ. Nucleotide sequence of the virulence gene virG of the Agrobacterium tumefaciens octopine Ti plasmid: significant homology between virG and the regulatory genes ompR, phoB and dye of E. coli. Nucleic Acids Res. 1986 Dec 22;14(24):9933–9942. [Europe PMC free article] [Abstract] [Google Scholar]
  • Mowbray SL, Foster DL, Koshland DE., Jr Proteolytic fragments identified with domains of the aspartate chemoreceptor. J Biol Chem. 1985 Sep 25;260(21):11711–11718. [Abstract] [Google Scholar]
  • Ninfa AJ, Magasanik B. Covalent modification of the glnG product, NRI, by the glnL product, NRII, regulates the transcription of the glnALG operon in Escherichia coli. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5909–5913. [Europe PMC free article] [Abstract] [Google Scholar]
  • Ordal GW. Bacterial chemotaxis: biochemistry of behavior in a single cell. Crit Rev Microbiol. 1985;12(2):95–130. [Abstract] [Google Scholar]
  • Ronson CW, Nixon BT, Ausubel FM. Conserved domains in bacterial regulatory proteins that respond to environmental stimuli. Cell. 1987 Jun 5;49(5):579–581. [Abstract] [Google Scholar]
  • Shaw CH, Ashby AM, Brown A, Royal C, Loake GJ, Shaw CH. virA and virG are the Ti-plasmid functions required for chemotaxis of Agrobacterium tumefaciens towards acetosyringone. Mol Microbiol. 1988 May;2(3):413–417. [Abstract] [Google Scholar]
  • Stachel SE, Zambryski PC. virA and virG control the plant-induced activation of the T-DNA transfer process of A. tumefaciens. Cell. 1986 Aug 1;46(3):325–333. [Abstract] [Google Scholar]
  • Strader CD, Sigal IS, Register RB, Candelore MR, Rands E, Dixon RA. Identification of residues required for ligand binding to the beta-adrenergic receptor. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4384–4388. [Europe PMC free article] [Abstract] [Google Scholar]
  • Vieira J, Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene. 1982 Oct;19(3):259–268. [Abstract] [Google Scholar]
  • Weber RF, Silverman PM. The cpx proteins of Escherichia coli K12. Structure of the cpxA polypeptide as an inner membrane component. J Mol Biol. 1988 Sep 20;203(2):467–478. [Abstract] [Google Scholar]
  • Winans SC, Ebert PR, Stachel SE, Gordon MP, Nester EW. A gene essential for Agrobacterium virulence is homologous to a family of positive regulatory loci. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8278–8282. [Europe PMC free article] [Abstract] [Google Scholar]
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