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Originally published In Press as doi:10.1074/jbc.M206573200 on August 21, 2002

J. Biol. Chem., Vol. 277, Issue 43, 41268-41273, October 25, 2002
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Identification of a Ras Palmitoyltransferase in Saccharomyces cerevisiae*

Sandra LoboDagger , Wendy K. Greentree§, Maurine E. Linder§, and Robert J. DeschenesDagger ||**

From the Dagger  Department of Biochemistry and the || Genetics Program, University of Iowa, Iowa City, Iowa 52240 and the § Department of Cell Biology and Physiology, Washington University, St. Louis, Missouri 63130

Most Ras proteins are posttranslationally modified by a palmitoyl lipid moiety through a thioester linkage. However, the mechanism by which this occurs is not known. Here, evidence is presented that the Ras2 protein of Saccharomyces cerevisiae is palmitoylated by a Ras protein acyltransferase (Ras PAT) encoded by the ERF2 and ERF4 genes. Erf2p is a 41-kDa protein localized to the membrane of the endoplasmic reticulum and contains a conserved DHHC cysteine-rich domain (DHHC-CRD). Erf2p co-purifies with Erf4p (26 kDa) when it is expressed in yeast or in Escherichia coli. The Erf2p/Erf4p complex is required for Ras PAT activity, and mutations within conserved residues (Cys189, His201, and Cys203) of the Erf2p DHHC-CRD domain abolish Ras PAT activity. Furthermore, a palmitoyl-Erf2p intermediate is detected suggesting that Erf2p is directly involved in palmitate transfer. ERF2 and ERF4 are the first genes identified that encode a palmitoyltransferase for a Ras GTPase.

* This work was supported by an American Heart Postdoctoral Fellowship (to S. L.), NCI, National Institutes of Health Grant CA50211(to R. J. D.), and National Institutes of Health Grant GM51466 (to M. E .L.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Established Investigator of the American Heart Association.

** To whom correspondence should be addressed: Dept. of Biochemistry, 3135 MERF, University of Iowa, Iowa City, IA 52242. Tel.: 319-335-7884; Fax: 319-334-4770; E-mail: Robert-deschenes@uiowa.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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