Nucleic Acids Research, 2003, Vol. 31, No. 6 e28
© 2003 Oxford University Press
Adaptation of the Ras-recruitment system to the analysis of interactions between membrane-associated proteins
Institut für Biologie III, Albert-Ludwigs-Universität, Schänzlestrasse 1, 79104 Freiburg, Germany
*To whom correspondence should be addressed. Tel: +49 761 203 2748; Fax: +49 761 203 2745; Email: fabian.koehler{at}biologie.uni-freiburg.de
Interactions of membrane-associated proteins play important roles in many cellular processes. The yeast two-hybrid assay is of limited utility for the analysis of such interactions, due to the need for soluble protein partners, whose interaction is assessed in the nucleus. The advent of the Ras-recruitment system (RRS) has enabled the study of membrane-associated proteins interacting with cytoplasmic proteins fused to Ras. Constitutive membrane association of the Ras fusion protein is expected to complement the growth defect of the yeast strain CDC25-2, assayed in the RRS, independent from the interaction with a membrane-bound partner. We describe the adaptation of the RRS to the analysis of interactions between two membrane-associated proteins using a model system. These results may facilitate the study of protein–protein interactions between membrane-bound proteins and further increase the utility of the RRS.