The yeast elongator histone acetylase requires Sit4-dependent dephosphorylation for toxin-target capacity

• Published in: Molecular biology of the cell Vol. 15; no. 3; pp. 1459 - 1469
• Main Authors: Jablonowski, Daniel, Fichtner, Lars, Stark, Michael J R, Schaffrath, Raffael
• Format: Journal Article
• Language: English
• Published: United States The American Society for Cell Biology 01.03.2004
• Subjects: Acetyltransferases - metabolism; Gene Expression Regulation, Fungal - drug effects; Histone Acetyltransferases; Killer Factors, Yeast; Kluyveromyces lactis; Mycotoxins - toxicity; Peptide Elongation Factors - metabolism; Phosphoprotein Phosphatases - metabolism; Phosphorylation - drug effects; Protein Binding - drug effects; Protein Phosphatase 2; Repressor Proteins - metabolism; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins - metabolism
• ISSN: 1059-1524; 1939-4586; 1059-1524
• DOI: 10.1091/mbc.E03-10-0750

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 sit4Delta-nuclei retain Tot4. Together with the findings that sit4Delta and totDelta cells phenocopy protection against zymocin and the ceramide-induced G1 block, Sit4 is functionally linked to Elongator in cell cycle events targetable by antizymotics.