Functional & Integrative Genomics

, Volume 1, Issue 6, pp 357–366

Two-hybrid-based analysis of protein–protein interactions of the yeast multidrug resistance protein, Pdr5p

  •  G. Subba Rao
  • Anand K. Bachhawat
  •  C. Gupta
Original Paper

DOI: 10.1007/s10142-001-0040-4

Cite this article as:
Subba Rao, G., Bachhawat, A.K. & Gupta, C. Funct Integr Genomics (2002) 1: 357. doi:10.1007/s10142-001-0040-4

Abstract.

The ATP-binding cassette (ABC) transporters are a large family of proteins responsible for the translocation of a variety of compounds across the membranes of both prokaryotes and eukaryotes. The inter-protein and intra-protein interactions in these traffic ATPases are still only poorly understood. In the present study we describe, for the first time, an extensive yeast two-hybrid (Y2H)-based analysis of the interactions of the cytoplasmic loops of the yeast pleiotropic drug resistance (Pdr) protein, Pdr5p, an ABC transporter of Saccharomycescerevisiae. Four of the major cytosolic loops that have been predicted for this protein [including the two nucleotide-binding domain (NBD)-containing loops and the cytosolic C-terminal region] were subjected to an extensive inter-domain interaction study in addition to being used as baits to identify potential interacting proteins within the cell using the Y2H system. Results of these studies have revealed that the first cytosolic loop (CL1) – containing the first NBD domain – and also the C-terminal region of Pdr5p interact with several candidate proteins. The possibility of an interaction between the CL1 loops of two neighboring Pdr5p molecules was also indicated, which could possibly have implications for dimerization of this protein.

Yeast two-hybrid system ABC transporter Pdr5p NBD1

Copyright information

© Springer-Verlag 2001

Authors and Affiliations

  •  G. Subba Rao
    • 1
  • Anand K. Bachhawat
    • 2
  •  C. Gupta
    • 1
  1. 1.Molecular and Structural Biology Division, Central Drug Research Institute, Chattar Manzil, Post Box 173, Lucknow 226001, India
  2. 2.Institute of Microbial Technology, Sector 39A, Chandigarh 160036, India