title: Identification of Pep4p as the protease responsible for formation of the SAGA-related SLIK protein complex.
abstract: The Saccharomyces cerevisiae Spt-Ada-Gcn5 acetyltransferase (SAGA) protein complex is a coactivator for transcription by RNA polymerase II and has various activities, including acetylation and deubuiqitination of histones and recruitment of TATA-binding protein to promoters. The Spt7p subunit is subject to proteolytic cleavage at its C terminus resulting in removal of the Spt8p-binding domain and generation of the SAGA-related SALSA/SAGA-like (SLIK) protein complex. Here, we report identification of the protease responsible for this cleavage. Screening of a protease knock-out collection revealed PEP4 to be required for cleavage of Spt7p within SAGA in vitro. Endogenous formation of truncated Spt7p was abolished in cells lacking PEP4. Purified Pep4p but not catalytic dead mutant Pep4p or unrelated Prc1p protease specifically cleaved Spt7p within SAGA into SLIK-related Spt7p. Interestingly, SAGA lacking Spt8p was more sensitive to Pep4p-mediated truncation of Spt7p, suggesting that Spt8p counteracted its own release from SAGA. Strains mimicking constitutive SLIK formation showed increased resistance to rapamycin treatment, suggesting a role for SLIK in regulating cellular responses to nutrient stress.
authors.affiliation: Department of Physiological Chemistry, University Medical Center Utrecht, Universiteitsweg 100, 3584 CG Utrecht, The Netherlands.
authors.forename: Gianpiero
authors.initials: G
authors.lastname: Spedale
authors.forename: Nikolai
authors.initials: N
authors.lastname: Mischerikow
authors.forename: Albert J R
authors.initials: AJ
authors.lastname: Heck
authors.forename: H T Marc
authors.initials: HT
authors.lastname: Timmers
authors.forename: W W M Pim
authors.initials: WW
authors.lastname: Pijnappel
mesh_terms: D016282:Aspartic Acid Endopeptidases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D025141:Drug Resistance, Fungal; D055786:Gene Knockout Techniques; D021122:Protein Subunits / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020123:Sirolimus / Q000494:pharmacology; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism*
chemical_list: D021122:Protein Subunits; C507785:SAGA complex, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; C499267:PEP4 protein, S cerevisiae; D016282:Aspartic Acid Endopeptidases; D020123:Sirolimus