The BioGRID | PUBLICATION VIEW

	Mol. Cell Apr (2007); 26(1):15-25
Mapping protein-protein interactions for the yeast ABC transporter Ycf1p by integrated split-ubiquitin membrane yeast two-hybrid analysis-
Paumi CM, Menendez J, Arnoldo A, Engels K, Iyer KR, Thaminy S, Georgiev O, Barral Y, Michaelis S, Stagljar I
Department of Cell Biology, The Johns Hopkins School of Medicine, Baltimore, MD 21205, USA-
Abstract: The ATP binding cassette -ABC- transporters are important in human health and disease and represent the largest family of transmembrane proteins; however, their highly hydrophobic nature complicates the use of standard biochemical approaches to identify interacting proteins- Here, we report the development of a modified version of the split-ubiquitin membrane yeast two-hybrid -MYTH- technology using genomically integrated "bait" constructs, hence the designation iMYTH- We used iMYTH in a library-screening format and identified six potential interacting partners of the yeast ABC transporter Ycf1p- Strains deleted for several of these genes result in arsenite sensitivity similar to a Deltaycf1 strain- Transport assays show that one of these, Tus1p, a guanine nucleotide exchange factor -GEF- for the small GTPase Rho1p, is a Rho1p-dependent-positive regulator of Ycf1p- Our study provides proof of principle that iMYTH is an ideal methodology to identify physiological interactors and regulators of ABC transporters and other yeast transmembrane proteins-
[PUBMED: 17434123]