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Published online December 15, 2008
doi:10.1083/jcb.200804003
The Journal of Cell Biology, Vol. 183, No. 6, 1061-1074
The Rockefeller University Press, 0021-9525 $30.00
© 2008 Baird et al.
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Article

Assembly of the PtdIns 4-kinase Stt4 complex at the plasma membrane requires Ypp1 and Efr3



Dan Baird1,2, Chris Stefan1,2, Anjon Audhya3, Sabine Weys1,2, and Scott D. Emr1,2

1 Weill Institute for Cell and Molecular Biology and 2 Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853
3 Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, WI 53706

Correspondence to Scott D. Emr: sde26{at}cornell.edu

The phosphoinositide phosphatidylinositol 4-phosphate (PtdIns4P) is an essential signaling lipid that regulates secretion and polarization of the actin cytoskeleton. In Saccharomyces cerevisiae, the PtdIns 4-kinase Stt4 catalyzes the synthesis of PtdIns4P at the plasma membrane (PM). In this paper, we identify and characterize two novel regulatory components of the Stt4 kinase complex, Ypp1 and Efr3. The essential gene YPP1 encodes a conserved protein that colocalizes with Stt4 at cortical punctate structures and regulates the stability of this lipid kinase. Accordingly, Ypp1 interacts with distinct regions on Stt4 that are necessary for the assembly and recruitment of multiple copies of the kinase into phosphoinositide kinase (PIK) patches. We identify the membrane protein Efr3 as an additional component of Stt4 PIK patches. Efr3 is essential for assembly of both Ypp1 and Stt4 at PIK patches. We conclude that Ypp1 and Efr3 are required for the formation and architecture of Stt4 PIK patches and ultimately PM-based PtdIns4P signaling.

D. Baird and C. Stefan contributed equally to this paper.

Abbreviations used in this paper: FPLC, fast protein liquid chromatography; PIP, phosphatidylinositol phosphate; PIK, phosphoinositide kinase; PM, plasma membrane; PtdIns, phosphatidylinositol; PtdIns4P, phosphatidylinositol 4-phosphate; RBO, rolling blackout; TPR, tetratricopeptide repeat.

© 2008 Baird et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).


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