Skip to main content

Yeast Two-Hybrid Protein-Protein Interaction Networks

  • Chapter
Proteomics and Protein-Protein Interactions

Part of the book series: Protein Reviews ((PRON,volume 3))

Abstract

The availability of complete genome sequences of numerous model organisms has initiated the development of new approaches in biological research to complement conventional biochemistry and genetics. Consequently, high-throughput methodologies also need to be applied in the emerging field of proteomics. Here, we discuss several methods that have been developed in the past years in order to characterize proteins and their functions on a large scale.We focus on the yeast two-hybrid system, which is the most widely used method to study protein-protein interactions and which has been used several times now to sucessfully map entire interaction networks on a large scale. We discuss small-scale pilot projects and how they have been upscaled to genome-wide screens, such as for the budding yeast Saccharomyces cerevisiae. We then compare the yeast two-hybrid system with several other screening methods that have been developed to investigate interactions between proteins in a high-throughput format, such as affinity purification methods coupled to mass spectrometry. Efficient adaptation of such methods to a high-throughput format, coupled with the increasing use of databases to compare interaction maps generated with different methods, will help in elucidating protein-protein interactions on a scale that would have been unthinkable just a few years ago.

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

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 169.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 219.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Aach, J., Bulyk, M.L., Church, G.M., Comander, J., Derti, A., Shendure, J. (2001). Computational comparison of two draft sequences of the human genome. Nature 409:856–859.

    Article  PubMed  CAS  Google Scholar 

  • Bartel, P.L., Roecklein, J.A., SenGupta, D., Fields, S. (1996). A protein linkage map of Escherichia coli bacteriophage T7. Nat. Genet. 12:72–77.

    Article  PubMed  CAS  Google Scholar 

  • Davy, A., Bello, P., Thierry-Mieg, N., Vaglio, P., Hitti, J., Doucette-Stamm, L., Thierry-Mieg, D., Reboul, J., Boulton, S., Walhout, A.J., Coux, O., Vidal, M. (2001). A protein-protein interaction map of the Caenorhabditis elegans 26S proteasome. EMBO Rep. 2:821–828.

    Article  PubMed  CAS  Google Scholar 

  • Fang, Y. and Macool, D.J. (2002). Development of a high-throughput yeast two-hybrid screening system to study protein-protein interactions in plants. Mol. Genet. Genomics 267:142–153.

    Article  PubMed  CAS  Google Scholar 

  • Dunn, J.J. and Studier, F.W. (1983). Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements. J. Mol. Biol. 166:477–535.

    Article  PubMed  CAS  Google Scholar 

  • Fields, S. and Song, O. (1989). A novel genetic system to detect protein-protein interactions. Nature 340:245–246.

    Article  PubMed  CAS  Google Scholar 

  • Finley, R.L., Jr. and Brent, R. (1994). Interaction mating reveals binary and ternary connections between Drosophila cell cycle regulators. Proc. Natl. Acad. Sci. USA 91:12980–12984.

    Article  PubMed  CAS  Google Scholar 

  • Flajolet, M., Rotondo, G., Daviet, L., Bergametti, F., Inchauspe, G., Tiollais, P., Transy, C., Legrain, P. (2000). A genomic approach of the hepatitis C virus generates a protein interaction map. Gene 242:369–379.

    Article  PubMed  CAS  Google Scholar 

  • Fromont-Racine, M., Rain, J.C., Legrain, P. (1997). Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens. Nat. Genet. 16:277–282.

    Article  PubMed  CAS  Google Scholar 

  • Gavin, A.C., Bosche, M., Krause, R., Grandi, P., Marzioch, M., Bauer, A., Schultz, J., Rick, J.M., Michon, A.M., Cruciat, C.M., Remor, M., Hofert, C., Schelder, M., Brajenovic, M., Ruffner, H., Merino, A., Klein, K., Hudak, M., Dickson, D., Rudi, T., Gnau, V., Bauch, A., Bastuck, S., Huhse, B., Leutwein, C., Heurtier, M.A., Copley, R.R., Edelmann, A., Querfurth, E., Rybin, V., Drewes, G., Raida, M., Bouwmeester, T., Bork, P., Seraphin, B., Kuster, B., Neubauer, G., and Superti-Furga, G. (2002). Functional organization of the yeast proteome by systematic analysis of protein complexes. Nature 415:141–147.

    Article  PubMed  CAS  Google Scholar 

  • Goffeau, A., Barrell, B., Bussey, H., Davis, R.W., Dujon, B., Feldmann, H., Galibert, F., Hoheisel, J.D., Jacq, C., Johnston, M., Louis, E.J., Mewes, H.W., Murakami, Y., Philippsen, P., Tettelin, H., Oliver, S.G. (1996). Life with 6000 genes. Science 274:563–567.

    Article  Google Scholar 

  • Ho, Y., Gruhler, A., Heilbut, A., Bader, G.D., Moore, L., Adams, S.L., Millar, A., Taylor, P., Bennett, K., Boutilier, K., Yang, L., Wolting, C., Donaldson, I., Schandorff, S., Shewnarane, J., Vo, M., Taggart, J., Goudreault, M., Muskat, B., Alfarano, C., Dewar, D., Lin, Z., Michalickova, K., Willems, A.R., Sassi, H., Nielsen, P.A., Rasmussen, K.J., Andersen, J.R., Johansen, L.E., Hansen, L.H., Jespersen, H., Podtelejnikov, A., Nielsen, E., Crawford, J., Poulsen, V., Sorensen, B.D., Matthiesen, J., Hendrickson, R.C., Gleeson, F., Pawson, T., Moran, M.F., Durocher, D., Mann, M., Hogue, C.W., Figeys, D., and Tyers, M. (2002). Systematic identification of protein complexes in Saccharomyces cerevisiae by mass spectrometry. Nature 415:180–183.

    Article  PubMed  CAS  Google Scholar 

  • Ito, T., Tashiro, K., Muta, S., Ozawa, R., Chiba, T., Nishizawa, M., Yamamoto, K. Kuhara, S., Sakaki, Y. (2000). Toward a protein-protein interaction map of the budding yeast: A comprehensive system to examine two-hybrid interactions in all possible combinations between the yeast proteins. Proc. Natl. Acad. Sci. USA 97:1143–1147.

    Article  PubMed  CAS  Google Scholar 

  • Ito, T., Chiba, T., Ozawa, R., Yoshida, M., Hattori, M., Sakaki, Y. (2001). A comprehensive two-hybrid analysis to explore the yeast protein interactome. Proc. Natl. Acad. Sci. USA 98:4569–4574.

    Article  PubMed  CAS  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  • Kumar, A. and Snyder, M. (2001) Emerging technologies in yeast genomics. Nat. Rev. Genet. 2:302–312.

    Article  PubMed  CAS  Google Scholar 

  • McCraith, S., Holtzman, T., Moss, B., Fields, S. (2000). Genome-wide analysis of vaccinia virus protein-protein interactions. Proc. Natl. Acad. Sci. USA 97:4879–4884.

    Article  PubMed  CAS  Google Scholar 

  • Mrowka, R., Patzak, A. (2001). Is there a bias in proteome research? Genome Res. 11:1971–1973.

    Article  PubMed  CAS  Google Scholar 

  • Rain, J.C., Selig, L., De Reuse, H., Battaglia, V., Reverdy, C., Simon, S., Lenzen, G., Petel, F., Wojcik, J., Schachter, V., Chemama, Y., Labigne, A., Legrain, P. (2001). The protein-protein interaction map of Helicobacter pylori. Nature 409:211–215.

    Article  PubMed  CAS  Google Scholar 

  • 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., Rothberg, J.M. (2000). A comprehensive analysis of protein-protein interactions in Saccharomyces cerevisiae. Nature 403:623–627.

    Article  PubMed  CAS  Google Scholar 

  • von Mering, C., Krause, R., et al. (2002). Comparative assessment of large-scale data sets of protein-protein interactions. Nature 417:399–403.

    Article  Google Scholar 

  • Walhout, A.J., Boulton, S.J., Vidal, M. (2000a). Yeast two-hybrid systems and protein interaction mapping projects for yeast and worm. Yeast 17:88–94.

    Article  PubMed  CAS  Google Scholar 

  • Walhout, A.J., Sordella, R., Lu, X., Hartley, J.L., Temple, G.F., Brasch, M.A., Thierry-Mieg, N., Vidal, M. (2000b). Protein interaction mapping in C. elegans using proteins involved in vulval development. Science 287:116–122.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2005 Springer Science+Business Media, Inc.

About this chapter

Cite this chapter

Auerbach, D., Stagljar, I. (2005). Yeast Two-Hybrid Protein-Protein Interaction Networks. In: Waksman, G. (eds) Proteomics and Protein-Protein Interactions. Protein Reviews, vol 3. Springer, Boston, MA. https://doi.org/10.1007/0-387-24532-4_2

Download citation

Publish with us

Policies and ethics