Submitted on November 28, 2004
Revised on December 30, 2004
Accepted on December 31, 2004
A genome wide screening approach for membrane-targeted proteins
Hanna Jaaro, Zehava Levy, and Mike Fainzilber
Department of Biological Chemistry, Weizmann Institute of Science, Rehovot 76100
Corresponding Author: mike.fainzilber{at}weizmann.ac.il
Membrane-associated proteins are critical for intra- and intercellular communication. Accordingly approaches are needed for rapid and comprehensive identification of all membrane-targeted gene products in a given cell or tissue. Here we describe a modification of the yeast Ras Recruitment System (RRS) to this end, and designate the modified approach the Ras Membrane Trap (RMT). A pilot RMT screen was carried out on the central nervous system of the mollusk Lymnaea stagnalis, a model organism from a phylum that still lacks a representative with a sequenced genome. 112 gene products were identified in the screen, of which 79 lack assignable homologues in available databases. Currently available annotation tools predicted membrane association of only 45% of the 112 proteins, although experimental verification in mammalian cells confirmed membrane association for all clones tested. Thus, genome annotation using currently available tools is likely to under-predict representation of membrane-associated gene products. The 32 proteins with known homologies include many targeted to the endoplasmic reticulum or the nucleus, thus RMT provides a tool that can cover intracellular membrane proteomes. Two sequences were found to represent gene families not found to date in invertebrate genomes, emphasizing the need for whole genome sequences from mollusks, and indeed from representatives of all major invertebrate phyla.