1887

Abstract

The development of a convenient and promising alternative to the various two-hybrid methods that are used to study protein–protein interactions is described. In this system, a lambdoid chimeric operator is recognized by a hybrid repressor formed by two chimeric monomers whose C-terminal domains are composed of heterologous proteins (or protein domains). Only if these proteins efficiently dimerize is a functional repressor formed able to bind the chimeric operator and shut off the synthesis of a downstream reporter gene. This new approach was tested with several interacting proteins ranging in size from less than 100 to more than 800 amino acids and, to date, no size or topology limit has been detected.

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2001-06-01
2019-12-13
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References

  1. Dente, L., Cesareni, G. & Cortese, R. ( 1983; ). pEMBL: a new family of single stranded plasmids. Nucleic Acids Res 11, 1645-1655.[CrossRef]
    [Google Scholar]
  2. Di Lallo, G., Anderluzzi, D., Ghelardini, P. & Paolozzi, L. ( 1999a; ). FtsZ dimerization in vivo. Mol Microbiol 2, 265-274.
    [Google Scholar]
  3. Di Lallo, G., Ghelardini, P. & Paolozzi, L. ( 1999b; ). Two-hybrid assay: construction of an Escherichia coli system to quantify homodimerization ability in vivo. Microbiology 145, 1485-1490.[CrossRef]
    [Google Scholar]
  4. Dove, S. L., Joung, J. K. & Hochschild, A. ( 1997; ). Activation of prokaryotic transcription through arbitrary protein–protein contacts. Nature 386, 627-630.[CrossRef]
    [Google Scholar]
  5. Fields, S. & Song, O. ( 1989; ). A novel genetic system to detect protein–protein interactions. Nature 340, 245-246.[CrossRef]
    [Google Scholar]
  6. Hanes, J. & Pluckthun, A. ( 1999; ). In vitro selection methods for screening of peptide and protein libraries. Curr Top Microbiol Immunol 243, 107-122.
    [Google Scholar]
  7. Hu, Z. & Lutkenhaus, J. ( 2000; ). Analysis of MinC reveals two independent domains involved in interaction with MinD and FtsZ. J Bacteriol 182, 3965-3971.[CrossRef]
    [Google Scholar]
  8. Hu, J. C., O’Shea, E. K., Kim, P. S. & Sauer, R. T. ( 1990; ). Sequence requirements for coiled-coils: analysis with λ repressor-GCN4 leucine zipper fusions. Science 250, 1400-1403.[CrossRef]
    [Google Scholar]
  9. Karimova, G., Pidoux, J., Ullmann, A. & Ladant, D. ( 1998; ). A bacterial two-hybrid system based on a reconstituted signal transduction pathway. Proc Natl Acad Sci USA 95, 5752-5756.[CrossRef]
    [Google Scholar]
  10. Karimova, G., Ullmann, A. & Ladant, D. ( 2000; ). A bacterial two-hybrid system that exploits a cAMP signaling cascade in Escherichia coli. Methods Enzymol 328, 59-73.
    [Google Scholar]
  11. Kornacker, M. G., Remsburg, B. & Menzel, R. ( 1998; ). Gene activation by the AraC protein can be inhibited by DNA looping between AraC and a LexA repressor that interacts with AraC: possible applications as a two-hybrid system. Mol Microbiol 30, 615-624.[CrossRef]
    [Google Scholar]
  12. Longo, F., Marchetti, M. A., Castagnoli, L., Battaglia, P. A. & Gigliani, F. ( 1995; ). A novel approach to protein-protein interaction: complex formation between the p53 tumour suppressor and the HIV Tat proteins. Biochem Biophys Res Commun 206, 326-334.[CrossRef]
    [Google Scholar]
  13. Miller, J. H. (1972). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  14. Pelletier, J. N., Arndt, K. M., Pluckthun, A. & Michnick, S. W. ( 1999; ). An in vivo library-versus-library selection of optimized protein-protein interactions. Nat Biotechnol 17, 683-690.[CrossRef]
    [Google Scholar]
  15. Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  16. Vidal, M. & Legrain, P. ( 1999; ). Yeast forward and reverse ‘n’-hybrid systems. Nucleic Acids Res 27, 919-929.[CrossRef]
    [Google Scholar]
  17. Wang, X., Huang, J., Mukherjee, A., Cao, C. & Lutkenhaus, J. ( 1997; ). Analysis of the interaction of FtsZ with itself, GTP, and FtsA. J Bacteriol 179, 5551-5559.
    [Google Scholar]
  18. Webster, C., Merryweather, C. & Brammar, W. ( 1992; ). Efficient repression by a heterodimeric repressor in Escherichia coli. Mol Microbiol 6, 371-377.[CrossRef]
    [Google Scholar]
  19. Wharton, R. P. & Ptashne, M. ( 1987; ). A new-specificity mutant of 434 repressor that defines an amino acid-base pair contact. Nature 326, 888-891.[CrossRef]
    [Google Scholar]
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