Palmitoylation of the vacuolar membrane protein Vac8p is essential for vacuole fusion in yeast (Veit, M., R. Laage, L. Dietrich, L. Wang, and C. Ungermann. 2001. EMBO J. 20:3145–3155; Wang, Y.X., E.J. Kauffman, J.E. Duex, and L.S. Weisman. 2001. J. Biol. Chem. 276:35133–35140). Proteins that contain an Asp-His-His-Cys (DHHC)–cysteine rich domain (CRD) are emerging as a family of protein acyltransferases, and are therefore candidates for mediators of Vac8p palmitoylation. Here we demonstrate that the DHHC-CRD proteins Pfa3p (protein fatty acyltransferase 3, encoded by YNL326c) and Swf1p are important for vacuole fusion. Cells lacking Pfa3p had fragmented vacuoles when stressed, and cells lacking both Pfa3p and Swf1p had fragmented vacuoles under normal growth conditions. Pfa3p promoted Vac8p membrane association and palmitoylation in vivo and partially purified Pfa3p palmitoylated Vac8p in vitro, establishing Vac8p as a substrate for palmitoylation by Pfa3p. Vac8p is the first N-myristoylated, palmitoylated protein identified as a substrate for a DHHC-CRD protein.
Skip Nav Destination
Article navigation
26 September 2005
Article|
September 26 2005
The vacuolar DHHC-CRD protein Pfa3p is a protein acyltransferase for Vac8p
Jessica E. Smotrys,
Jessica E. Smotrys
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110
Search for other works by this author on:
Marissa J. Schoenfish,
Marissa J. Schoenfish
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110
Search for other works by this author on:
Monica A. Stutz,
Monica A. Stutz
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110
Search for other works by this author on:
Maurine E. Linder
Maurine E. Linder
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110
Search for other works by this author on:
Jessica E. Smotrys
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110
Marissa J. Schoenfish
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110
Monica A. Stutz
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110
Maurine E. Linder
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110
Correspondence to M.E. Linder: [email protected]
J.E. Smotrys and M.J. Schoenfish contributed equally to this work.
Abbreviations used in this paper: DHHC-CRD, Asp-His-His-Cys cysteine-rich domain; LB, lysis buffer; myr-Vac8p, N-myristoylated Vac8p; NF, nonfluorescent media; palm-CoA, palmitoyl-CoenzymeA; PAT, protein acyltransferase; PFA, protein fatty acyltransferase; WT, wild type.
Received:
July 12 2005
Accepted:
August 19 2005
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2005
J Cell Biol (2005) 170 (7): 1091–1099.
Article history
Received:
July 12 2005
Accepted:
August 19 2005
Citation
Jessica E. Smotrys, Marissa J. Schoenfish, Monica A. Stutz, Maurine E. Linder; The vacuolar DHHC-CRD protein Pfa3p is a protein acyltransferase for Vac8p . J Cell Biol 26 September 2005; 170 (7): 1091–1099. doi: https://doi.org/10.1083/jcb.200507048
Download citation file:
Sign in
Don't already have an account? Register
Client Account
You could not be signed in. Please check your email address / username and password and try again.
Could not validate captcha. Please try again.
Sign in via your Institution
Sign in via your InstitutionEmail alerts
Advertisement
Advertisement