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Originally published as MBC in Press, 10.1091/mbc.E03-10-0750 on January 12, 2004

Vol. 15, Issue 3, 1459-1469, March 2004

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The Yeast Elongator Histone Acetylase Requires Sit4-dependent Dephosphorylation for Toxin-Target Capacity

Daniel Jablonowski *, Lars Fichtner *, Michael J.R. Stark {dagger}, and Raffael Schaffrath * {ddagger}

* Biologicum, Institut für Genetik, Martin-Luther-Universität Halle-Wittenberg, D-06120 Halle, Saale, Germany; {dagger} Division of Gene Regulation and Expression, School of Life Sciences, University of Dundee, MSI/WTB Complex, Dundee DD1 5EH, United Kingdom

Submitted October 21, 2003; Revised November 29, 2003; Accepted December 4, 2003
Monitoring Editor: Trisha Davis

Kluyveromyces lactis zymocin, a heterotrimeric toxin complex, imposes a G1 cell cycle block on Saccharomyces cerevisiae that requires the toxin-target (TOT) function of holo-Elongator, a six-subunit histone acetylase. Here, we demonstrate that Elongator is a phospho-complex. Phosphorylation of its largest subunit Tot1 (Elp1) is supported by Kti11, an Elongator-interactor essential for zymocin action. Tot1 dephosphorylation depends on the Sit4 phosphatase and its associators Sap185 and Sap190. Zymocin-resistant cells lacking or overproducing Elongator-associator Tot4 (Kti12), respectively, abolish or intensify Tot1 phosphorylation. Excess Sit4·Sap190 antagonizes the latter scenario to reinstate zymocin sensitivity in multicopy TOT4 cells, suggesting physical competition between Sit4 and Tot4. Consistently, Sit4 and Tot4 mutually oppose Tot1 de-/phosphorylation, which is dispensable for integrity of holo-Elongator but crucial for the TOT-dependent G1 block by zymocin. Moreover, Sit4, Tot4, and Tot1 cofractionate, Sit4 is nucleocytoplasmically localized, and sit4{Delta}-nuclei retain Tot4. Together with the findings that sit4{Delta} and tot{Delta} cells phenocopy protection against zymocin and the ceramide-induced G1 block, Sit4 is functionally linked to Elongator in cell cycle events targetable by antizymotics.

Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E03-10-0750. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E03-10-0750.

{ddagger} Corresponding author. E-mail address: schaffrath{at}genetik.unihalle.de.




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