Skip to main content
SpringerLink
Log in

Identification of a S-ribonuclease-binding protein in Petunia hybrida

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

To investigate protein-protein interactions in gametophytic self-incompatibility, we used a yeast two-hybrid assay to identify proteins that could interact with the S-ribonuclease protein. These assays identified a pollen-expressed protein, which we have named PhSBP1, that appears to bind with a high degree of specificity to the Petunia hybrida S-ribonuclease. Although PhSBP1 activates reporter gene expression only when expressed in tandem with a S-RNAse bait protein, binding is not allele-specific. Sequence analysis demonstrated that PhSBP1 contained a C-terminal cysteine-rich region that includes a RING-HC domain. Because many RING-finger domain proteins appear to function as E3 ubiquitin ligases, our results suggest that ubiquitination and protein degradation may play a role in regulating self-incompatibility interactions. Together, these results suggest that PhSBP1 may be a candidate for the recently proposed general inhibitor (RI) of self-incompatibility ribonucleases.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Adams, A., Gottschling, D.E., Kaiser, C.A. and Stearns T. 1997. Methods in Yeast Genetics. Cold Spring Harbor Laboratory Press, Plainview, NY.

    Google Scholar 

  • Ai, Y., Singh, A., Coleman, C.E., Ioerger, T.R., Kheyr-Pour, A. and Kao, T.-H. 1990. Self-incompatibility in Petunia in-flata: isolation and characterization of cDNAs encoding three S-allele-associated proteins. Sex. Plant Reprod. 3: 130–138.

    Google Scholar 

  • Ai, Y., Kron, E. and Kao, T.-H. 1991. S-alleles are retained and expressed in a self-compatible cultivar of Petunia hybrida. Mol. Gen. Genet. 230: 353–358.

    Google Scholar 

  • Alché, J.D., Butowt, R., Castro, A.J. and Rodríguez-García, M.I. 2000. Ubiquitin and ubiquitin-conjugated proteins in the olive (Olea europaea L.) pollen. Sex. Plant Reprod. 12: 285–291.

    Google Scholar 

  • Altschul, S.F., Madden, T.L., Schäffer, A.A., Zhang, J., Zhang, Z., Miller, W. and Lipman, D.J. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl. Acids Res. 25: 3389–3402.

    Google Scholar 

  • Anderson, M.A., McFadden, G.I., Bernatsky, R., Atkinson, A., Orpin, T., Dedman, H., Tregear, G., Fernley, R. and Clarke, A.E. 1989. Sequence variability of three alleles of the self-incompatibility gene of Nicotiana alata. Plant Cell 1: 483–491.

    Google Scholar 

  • Appel, R.D., Bairoch, A. and Hochstrasser, D.F. 1994. A new generation of information retrieval tools for biologists: the example of the ExPASy WWWserver. Trends Biochem. Sci. 19: 258–260.

    Google Scholar 

  • Barlow, P.N., Luisi, B., Milner, A., Elliott, M. and Everett, R. 1994. Structure of the C3HC4 domain by 1H-nuclear magnetic resonance spectroscopy: a new structural class of zinc finger. J. Mol. Biol. 237: 201–211.

    Google Scholar 

  • Bendixen, C., Gangloff, S. and Rothstein, R. 1994. A yeast mating-selection scheme for detection of protein-protein interactions. Nucl. Acids Res. 22: 1778–1779.

    Google Scholar 

  • Beuken, E., Vink, C. and Bruggeman, C.A. 1998. One-step procedure for screening recombinant plasmids by size. Biotechniques 24: 748–750.

    Google Scholar 

  • Borden, K.L.B. and Freemont, P.S. 1996. The RING finger domain: a recent example of a sequence-structure family. Curr. Opin. Struct. Biol. 6: 395–401.

    Google Scholar 

  • Callis, J. and Bedinger, P. 1994. Developmentally regulated loss of ubiquitin and ubiquitinated proteins during pollen maturation in maize. Proc. Natl. Acad. Sci. USA 91: 6074–6077.

    Google Scholar 

  • Chawla, B., Bernatzky, R., Liang, W. and Marcotrigiano, M. 1997. Breakdown of self-incompatibility in tetraploid Lycopersicon peruvianum: inheritance and expression of S-related proteins Theor. Appl. Genet. 95: 992–996.

    Google Scholar 

  • Chien, C.-T., Bartel, P.L., Sternglanz, R. and Fields, S. 1991. The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc. Natl. Acad. Sci. USA 88: 9578–9582.

    Google Scholar 

  • Clark, K.R. and Sims, T.L. 1994. The S-ribonuclease gene of Petunia hybrida is expressed in nonstylar tissue, including immature anthers. Plant Physiol. 106: 25–36.

    Google Scholar 

  • Clark, K.R., Okuley, J.J., Collins, P.D. and Sims, T.L. 1990. Sequence variability and developmental expression of S-alleles in self-incompatible and pseudo-self-compatible petunia. Plant Cell 2: 815–826.

    Google Scholar 

  • Clontech. 1998. Matchmaker Gal4 two-hybrid user manual (PT3061-1).

  • Dana, M.N. and Ascher, P.D. 1986. Sexually localized expressin of pseudo-self compatibility (PSC) in Petunia ×hybrida Hort. Theor. Appl. Genet. 71: 578–584.

    Google Scholar 

  • Dean, A.D, and Greenwald, J.E. 1995. Use of filtered pipette tips to elute DNA from agarose gels. BioTechniques 18: 980.

    Google Scholar 

  • de Nettancourt, D. 1977. Incompatibility in angiosperms. In: R. Frankel, G.A.E. Gall, M. Grossman and H.F. Linskens (Eds.) Monographs on Theoretical and Applied Genetics, Vol. 3, Springer-Verlag, Berlin.

    Google Scholar 

  • Dodds, P.N., Bonig., I., Du., H., Rödin., J., Anderson., M.A., Newbigin, E. and Clarke, A.E. 1993. S-RNase gene of Nicotiana alata is expressed in developing pollen. Plant Cell 5: 1771–1782.

    Google Scholar 

  • Doughty, J., Dixon, S., Hiscock, S.J., Willis, A.C., Parkin, I.A.P. and Dickinson, H.G. 1998. PCP-A1, a defensin-like Brassica pollen coat protein that binds the S locus glycoprotein, is the product of gametophytic gene expression. Plant Cell 10: 1333–1347.

    Google Scholar 

  • Doughty, J., Wong, H.Y. and Dickinson, H.G. 2000. Cysteine-rich pollen coat proteins (PCPs) and their interactions with stigmatic S (incompatibility) and S-related proteins in Brassica: putative roles in SI and pollination. Ann. Bot. 85A: 161–169.

    Google Scholar 

  • Fields, S. and Sternglanz, R. 1994. The two-hybrid system: an assay for protein-protein interactions. Trends Genet. 10: 286–292.

    Google Scholar 

  • Flaschenreim, D.R. and Ascher, P.D. 1979. S allele discrimination in styles of Petunia hybrida bearing stylar-conditioned pseudo-self compatibility. Theor. Appl. Genet. 55: 23–28.

    Google Scholar 

  • Freemont, P.S. 2000. Ubiquitination: RING for destruction? Curr. Biol. 10: R84–R87.

    Google Scholar 

  • Garnier, J., Gibrat, J.-F. and Robson, B. 1996. GOR secondary structure prediction method version IV. Meth. Enzymol. 266: 540–553.

    Google Scholar 

  • Golz, J.F., Su, V., Clarke, A.E. and Newbigin, E. 1999.A molecular description of mutations affecting the pollen component of the Nicotiana alata S-locus. Genetics 152: 1123–1135.

    Google Scholar 

  • Golz, J.F., Clarke, A.E. and Newbigin, E. 2000. Mutational approaches to the study of self-incompatibility: revisiting the pollen-part mutants. Ann. Bot. 85A: 95–103.

    Google Scholar 

  • Gu, T., Mazzurco, M., Sulaman, W. Matias, D.D. and Goring, D.R. 1998. Binding of an arm repeat protein to the kinase domain of the S-locus receptor kinase. Proc. Natl. Acad. Sci. USA 95: 382–387.

    Google Scholar 

  • Hearn, M.J., Franklin, F.C.H. and Ride, J.P. 1996. Identification of a membrane glycoprotein in pollen of Papaver rhoeas which binds stigmatic self-incompatibility (S-) proteins. Plant J. 9: 467–475.

    Google Scholar 

  • Huang, S., Lee, H.-S., Karunanandaa, B. and Kao, T.-H. 1994. Ribonuclease activity of Petunia inflata S proteins is essential for rejection of self-pollen. Plant Cell 6: 1021–1028.

    Google Scholar 

  • Ioerger, T.R., Gohlke, J.R., Xu, B. and Kao, T.-H. 1991. Primary structural features of the self-incompatibility protein in Solanaceae. Sex. Plant Reprod. 4: 81–87.

    Google Scholar 

  • Jackson, P.K., Eldridge, A.G., Freed, E., Furstenthal, L., Hsu, J.Y., Kaiser, B.K. and Reimann, J.D.R. 2000. The lore of the RINGs: substrate recognition and catalysis by ubiquitin ligases. Trends Cell Biol. 10: 429–439.

    Google Scholar 

  • Joazeiro, C.A.P. and Weissmann, A.M. 2000. RING finger proteins: mediators of ubiquitin ligase activity. Cell102: 549–552.

    Google Scholar 

  • James, P., Halladay, J. and Craig, E.A. 1996. Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics 144: 1425–1436.

    Google Scholar 

  • James., P. 1997. PJ69-4A Update.

  • Jordan, N.D., Kakeda, K., Conner, A., Ride, J.P., Franklin-Tong, V.E. and Franklin, F.C.H. 1999. S-protein mutants indicate a functional role for SBP in the self-incompatibility reaction of Papaver rhoeas. Plant J. 20: 119–125.

    Google Scholar 

  • Lecrenier, N., Foury, F. and Goffeau, A. 1998. Two-hybrid systematic screening of the yeast proteome. BioEssays 20: 1–6.

    Google Scholar 

  • Lee, H.-S., Huang, S. and Kao, T.-H. 1994. S-proteins control rejection of incompatible pollen in Petunia inflata. Nature 367: 560–563.

    Google Scholar 

  • Lewis, D. 1961. Chromosome fragments and mutation of the incompatibility gene. Nature 190: 990–991.

    Google Scholar 

  • Lorick, K.L., Jensen, J.P., Fang, S., Ong, O.M., Hatakeyama, S. and Weissman, A.M. 1999 RING fingers mediate ubiquitin-conjugating enzyme (E2)-dependent ubiquitination. Proc. Natl. Acad. Sci. USA 96: 11364–11369.

    Google Scholar 

  • Luu, D.-T., Qin, X., Morse, D. and Cappadocia, M. 2000. S-RNase uptake by compatible pollen tubes in gametophytic self-incompatibility. Nature 407: 649–651.

    Google Scholar 

  • Matton, D.P., Maes, C., Laublin, G., Qin, X.K., Bertrand, C., Morse, D. and Cappadocia, M. 1997. Hypervariable domains of self-incompatibility RNases mediate allele-specific pollen recognition. Plant Cell 9: 1757–1766.

    Google Scholar 

  • Matton, D.P., Luu, D.T., Xike, Q., Laublin, G., O'Brien, M., Maes, O., Morse, D. and Cappadocia, M. 1999. Production of an S-RNase with dual specificity suggests a novel hypothesis for the generation of new S alleles. Plant Cell 11: 2087–2097.

    Google Scholar 

  • McAlister-Henn, L., Gibson, N. and Panisko, E. 1999. Applications of the yeast two-hybrid system. Methods 19: 330–337.

    Google Scholar 

  • McLure, B., Mou, B., Canevascini, S. and Bernatzky, R. 1999.A small asparagine-rich protein required for S-allele-specific pollen rejection in Nicotiana. Proc. Natl. Acad. Sci. 96: 13548–13553.

    Google Scholar 

  • McClure, B.A., Haring, V., Ebert, P.R., Anderson, M.A., Simp-son, R.J., Sakiyama, F. and Clarke A.E. 1989. Style self-incompatibility gene products of Nicotiana alata are ribonucleases. Nature 342: 955–957.

    Google Scholar 

  • McClure, B.A., Gray, J.E., Anderson, M.A. and Clarke, A.E. 1990. Self-incompatibility in Nicotiana alata involves degradation of pollen rRNA. Nature 347: 757–760.

    Google Scholar 

  • McCubbin, A.G., Chung, Y.Y. and Kao, T.-H. 1997. A mutant S 3 Rnase of Petunia inflata lacking RNase activity has a dominant-negative effect on self-incompatibility interactions. Plant Cell 9: 85–95.

    Google Scholar 

  • Murfett, J., Atherton, T.L., Mou, B., Gasser, C.S. and McClure, B.A. 1994. S-RNase expressed in transgenic Nicotiana causes S-allele-specific pollen rejection. Nature 367: 563–566.

    Google Scholar 

  • Pandey, K. 1967. Elements of the S-gene complex II. Mutations and complementation at the SI locus in Nicotiana alata. Heredity 22: 255–284.

    Google Scholar 

  • Parry, S., Newbigin, E., Craik, D., Nakamura, K.T., Bacic, A. and Oxley, D. 1998. Structural analysis and molecular model of a self-incompatibility RNase from wild tomato. Plant Physiol. 116: 463–469.

    Google Scholar 

  • Promega Corporation. 1999. Riboprobe® In Vitro Transcription Systems, Technical Manual 016.16

  • Rost, B. and Sander, C. 1993. Prediction of protein secondary structure at better than 70% accuracy. J. Mol. Biol. 232: 584–599.

    Google Scholar 

  • Rost, B. and Sander, C. 1994. Combining evolutionary information and neural networks to predict protein secondary structure. Proteins 19: 55–77.

    Google Scholar 

  • Royo, J., Kunz, C., Kowyama, Y., Anderson, M., Clarke, A.E. and Newbigin, E. 1994. Loss of a histidine residue at the active site of S-locus ribonuclease is associated with self-compatibility in Lycopersicon peruvianum. Proc. Natl. Acad. Sci. USA 91: 6511–6514.

    Google Scholar 

  • Sambrook, J., Fritsch, E.F. and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Plainview, NY.

    Google Scholar 

  • Saurin, A.J., Borden, K.L.B., Boddy, M.N. and Freemont, P.S. 1996. Does this have a familiar RING? Trends Biochem. Sci. 21: 208–214.

    Google Scholar 

  • Sims, T.L. 1993. Genetic regulation of self-incompatibility. CRC Crit. Rev. Plant Sci. 12: 129–167.

    Google Scholar 

  • Sims, T.L. 1994. Molecular genetics of gametophytic self-incompatibility in Petunia hybrida. In: E.G. Williams, A.E. Clarke and R.B. Knox (Eds.) Genetic Control of Self-Incompatibility and Reproductive Development in Flowering Plants, Kluwer Academic Publishers, Dordrecht, Netherlands, pp. 19–41.

    Google Scholar 

  • Schopfer, C.R., Nasrallah, M.E. and Nasrallah, J.B. 1999. The male determinant of self-incompatibility in Brassica. Science 286: 1697–1700.

    Google Scholar 

  • Solovyev, V.V. and Salamov, A.A. 1994. Predicting á-helix and β-strand segments of globular proteins. CABIOS 10: 661–669.

    Google Scholar 

  • Thompson, J.D., Higgins, D.G. and Gibson, T.J. 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucle. Acids Res. 22: 4673–4680.

    Google Scholar 

  • Twell, D., Wing, R., Yamaguchi, J. and McCormick, S. 1989. Isolation and expression of an anther-specific gene from tomato. Mol. Gen. Genet. 217: 240–245.

    Google Scholar 

  • Twell, D., Yamaguchi, J. and McCormick, S. 1990. Pollen-specific gene expression in transgenic plants: coordinate regulation of two different tomato gene promoters during microsporogenesis. Development 109: 705–713.

    Google Scholar 

  • Twell, D., Yamaguchi, J., Wing, R.A., Ushiba, J. and McCormick, S. 1991. Promoter analysis of genes that are coordinately expressed during pollen development reveals pollen-specific enhancer sequences and shared regulatory elements. Genes Dev. 5: 496–507.

    Google Scholar 

  • Tyers, M. and Jorgensen, P. 2000. Proteolysis and the cell cycle: with this RING I do thee destroy. Curr. Opin. Genet. Dev. 10: 54–64.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biological Sciences and Plant Molecular Biology Center, Northern Illinois University, DeKalb, IL, 60115-2861, USA

    Thomas L. Sims & Maja Ordanic

Authors
  1. Thomas L. Sims
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maja Ordanic
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sims, T.L., Ordanic, M. Identification of a S-ribonuclease-binding protein in Petunia hybrida . Plant Mol Biol 47, 771–783 (2001). https://doi.org/10.1023/A:1013639528858

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1013639528858

Search

Navigation