Abstract
Plant research is moving into the post-genomic era. Proteomic-based strategies are now being developed to study functional aspects of the genes predicted from the various genome-sequencing initiatives. All biological processes depend on interactions formed between proteins and the mapping of such interactions on a global scale is providing interesting functional insights. One of the techniques that has proved itself invaluable in the mapping of protein-protein interactions is the yeast two-hybrid system. This system is a sensitive molecular genetic approach for studying protein-protein interactions in vivo. In this review we will introduce the yeast two-hybrid system, discuss modifications of the system that may be of interest to the plant science community and suggest potential applications of the technology.
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References
Aronheim, A., Zandi, E., Hennemann, H., Elledge, S.J. and Karin, M. 1997. Isolation of an AP-1 repressor by a novel method for detecting protein-protein interactions. Mol. Cell Biol. 17: 3094–3102.
Brent, R. and Finley, R.L. 1997. Understanding gene and allele function with two-hybrid methods. Annu. Rev. Genet. 31: 663–704.
Brent, R. and Ptashne, M. 1984. A bacterial repressor protein or a yeast transcriptional terminator can block upstream activation of a yeast gene. Nature 312: 612–615
Broder, Y.C., Katz, S. and Aronheim, A. 1998. The Ras recruitment system, a novel approach to the study of protein-protein interactions. Curr. Biol. 8: 1121–1124.
Causier, B., Weir, I. and Davies, B. 1999. MADS-box factors in hermaphrodite flower development. In: C.C. Ainsworth (Ed.) Sex Determination in Plants, Bios Scientific Publishers, Oxford, pp. 1–23.
Colas, P. and Brent, R. 1998. The impact of two-hybrid and related methods on biotechnology. Trends Biotechnol. 16: 355–363.
Davies, B., Egea-Cortines, M., de Andrade Silva, E., Saedler, H. and Sommer, H. 1996. Multiple interactions amongst floral homeotic proteins. EMBO J. 15: 4330–4343.
Dent, C.L., Smith, M.D. and Latchman, D.S. 1999. The DNA mobility shift assay. In: D.S. Latchman (Ed.) Transcription Factors: A Practical Approach, Oxford University Press, Oxford, pp. 1–25.
Egea-Cortines, M., Saedler, H. and Sommer, H. 1999. Ternary complex formation between the MADS-box proteins SQUAMOSA, DEFICIENS and GLOBOSA is involved in the control of floral architecture in Antirrhinum majus. EMBO J. 18: 5370–5379.
Ellis et al.-this issue
Fashena, S.J., Serebriiskii, I. and Golemis, E.A. 2000. The continued evolution of two-hybrid screening approaches in yeast: how to outwit different preys with different baits. Gene 250: 1–14.
Fields, S. and Song, O.-K. 1989. A novel genetic system to detect protein-protein interactions. Nature 340: 245–246.
Gietz, R.D. and Schiestl, R.H. 1995. Transforming yeast with DNA. Meths Mol Cell Biol 5: 255–269.
Gyuris, J., Golemis, E., Chertov, H. and Brent, R. 1993. Cdi1, a human G1 and S phase protein phosphatase that associates with Cdk2. Cell 75: 791–803.
Honma, T. and Goto, K. 2001. Complexes of MADS-box proteins are sufficient to convert leaves into floral organs. Nature 409: 525–529.
Jarillo, J.A., Capel, J., Tang, R.H., Yang, H.Q., Alonso, J.M., Ecker, J.R. and Cashmore, A.R. 2001. An Arabidopsis circadian clock component interacts with both CRY1 and phyB. Nature 410: 487–490.
Johnsson, N. and Varshavsky, A. 1994. Split ubiquitin as a sensor of protein interactions in vivo. Proc. Natl. Acad. Sci. USA 91: 10340–10344.
Joung, J.K., Ramm, E.I. and Pabo, C.O. 2000. A bacterial twohybrid selection system for studying protein-DNA and proteinprotein interactions. Proc. Natl. Acad. Sci. USA 97: 7382–7387.
Leanna, C.A. and Hannink, M. 1996. The reverse two-hybrid system: a genetic scheme for selection against specific protein/ protein interactions. Nucl. Acids Res. 24: 3341–3347.
Long, J.A., Moan, E.I., Medford, J. and Barton, M.K. 1996. A member of the KNOTTED class of homeodomain proteins encoded by the SHOOTMERISTEMLESS gene of Arabidopsis. Nature 379: 66–69.
Luo, Y., Batalao, A., Zhou, H. and Zhu, L. 1997. Mammalian two-hybrid system. A complementary approach to the yeast two-hybrid system. Biotechniques 22: 350–352.
Mazzurco, M., Sulaman,W., Elina, H., Cock, J.M. and Goring, D.R. 2001. Further analysis of the interactions between the Brassica S receptor kinase and three interacting proteins (ARC1, THL1 and THL2) in the yeast two-hybrid system. Plant Mol. Biol. 45: 365–376.
Ouellet, F., Overvoorde, P.J. and Theologis, A. 2001. IAA17/AXR3: Biochemical insight into an auxin mutant phenotype. Plant Cell 13: 829–841.
Pelaz, S., Ditta, G.S., Baumann, E., Wisman, E. and Yanofsky, M.F. 2000. B and C floral organ identity functions require SEPALLATA MADS-box genes. Nature 405: 200–203.
Rain, J.-C., Selig, L., De Reuse, H., Battaglia, V., Reverdy, C., Simon, S., Lenzen, G., Petel, F., Wojcik, J., Schächter, V., Chemama, V.Y., Labigne, A. and Legrain, P. 2001. The protein-protein interaction map of Helicobacter pylori. Nature 409: 211–215.
Serebriiskii, I., Khazak, V. and Gomelis, E.A. 1999. A twohybrid dual bait system to discriminate specificity of protein interactions. J. Biol. Chem. 274: 17080–17087.
Serebriiskii, I.G., Toby, G.G. and Golemis, E.A. 2000a. Two-hybrid system for the characterization of protein-protein interactions in E. coli. Biotechniques 29: 288–296.
Serebriiskii, I.G., Estojak, J., Berman, M. and Golemis, E.A. 2000b. Approaches to detecting false positives in yeast two-hybrid systems. Biotechniques 28: 328–336.
Stagljar, I., Korostensky, C., Johnsson, N. and Te Heesen, S. 1998. A genetic system based on split-ubiquitin for the analysis of interactions between membrane proteins in vivo. Proc. Natl. Acad. Sci. USA 95: 5187–5192.
Uetz, P., Giot, L., Cagney, G., Mansfield, T.A., Judson, R.S., Knight, J.R., Lockshon, D., Narayan, V., Srinivasan, M., Pochart, P., Qureshi-Emili, A., Li, Y., Godwin, B., Conover, D., Kalbfleisch, T., Vijayadamodar, G., Yang, M., Johnston, M., Fields, S. and Rothberg, J.M. 2000. A comprehensive analysis of protein-protein interactions in Saccharomyces cerevisiae. Nature 403: 623–627.
Vidal, M., Brachmann, R.K., Fattaey, A., Harlow, E. and Boeke, J.D. 1996a. Reverse two-hybrid and one-hybrid systems to detect dissociation of protein-protein and DNA-protein interactions. Proc. Natl. Acad. Sci. USA 93: 10315–10320.
Vidal, M., Braun, P., Chen, E., Boeke, J.D. and Harlow, E. 1996b. Genetic characterization of a mammalian protein-protein interaction domain by using a yeast reverse two-hybrid system. Proc. Natl. Acad. Sci. USA 93: 10321–10326.
Visser-this issue
Vitale-this issue
Walhout, A.J.M., Sordella, R., Lu, X., Hartley, J.L., Temple, G.F., Brasch, M.A., Thierry-Mieg, N. and Vidal, M. 2000. Protein interaction mapping in C. elegans using proteins involved in vulval development. Science 287: 116–122.
Zhang, J. and Lautar, S. 1996. A yeast three-hybrid method to clone ternary protein complex components. Anal. Biochem. 242: 68–72.
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Causier, B., Davies, B. Analysing protein-protein interactions with the yeast two-hybrid system. Plant Mol Biol 50, 855–870 (2002). https://doi.org/10.1023/A:1021214007897
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DOI: https://doi.org/10.1023/A:1021214007897