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Genome‐wide lethality screen identifies new PI4,5P 2 effectors that regulate the actin cytoskeleton
Author(s) -
Audhya Anjon,
Loewith Robbie,
Parsons Ainslie B,
Gao Lu,
Tabuchi Mitsuaki,
Zhou Huilin,
Boone Charles,
Hall Michael N,
Emr Scott D
Publication year - 2004
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1038/sj.emboj.7600384
Subject(s) - biology , actin cytoskeleton , microbiology and biotechnology , cytoskeleton , effector , actin , kinase , genetics , cell
To further understand the roles played by the essential phosphoinositide PI4,5P 2 , we have used a synthetic lethal analysis, which systematically combined the mss4 ts mutation, partially defective in PI4P 5‐kinase activity, with each of approximately 4700 deletion mutations. This genomic screening technique uncovered numerous new candidate effectors and regulators of PI4,5P 2 in yeast. In particular, we identified Slm1 (Yil105c), a previously uncharacterized PI4,5P 2 binding protein. Like Mss4, Slm1 and its homolog Slm2 (Ynl047c) were required for actin cytoskeleton polarization and viability. Co‐immunoprecipitation experiments revealed that Slm1 interacts with a component of TORC2, a Tor2 kinase‐containing complex, which also regulates the actin cytoskeleton. Consistent with these findings, phosphorylation of Slm1 and Slm2 was dependent on TORC2 protein kinase activity, both in vivo and in vitro , and Slm1 localization required both PI4,5P 2 and functional TORC2. Together, these data suggest that Slm1 and Slm2 function downstream of PI4,5P 2 and the TORC2 kinase pathway to control actin cytoskeleton organization.