Stable incorporation of sequence specific repressors Ash1 and Ume6 into the Rpd3L complex.
Histone deacetylation by Saccharomyces cerevisiae Rpd3 represses genes regulated by the Ash1 and Ume6 DNA-binding proteins. Rpd3 exists in a small 0.6 MDa (Rpd3S) and large 1.2 MDa (Rpd3L) corepressor complex. In this report, we identify by mass spectrometry and MudPIT the subunits of the Rpd3L complex. These included Rpd3, Sds3, Pho23, Dep1, Rxt2, Sin3, Ash1, Ume1, Sap30, Cti6, Rxt3 and Ume6. Dep1 and Sds3, unique components of Rpd3L, were required for Rpd3L integrity and HDAC activity. Similar to RPD3, deletion of DEP1 enhanced telomeric silencing and derepressed INO1. Two sequence-specific repressors, Ash1 and Ume6, were stably associated with Rpd3L. While both of these proteins localized to the INO1 and HO promoters, the repression of these genes were dependent only on Ume6 and Ash1, respectively. Thus, the Rpd3L complex is directly recruited to specific promoters through multiple integral DNA-binding proteins.
Carrozza
Michael J
MJ
Stowers Institute for Medical Research1000 East 50th Street, Kansas City, MO 64110, USA.
Florens
Laurence
L
Swanson
Selene K
SK
Shia
Wei-Jong
WJ
Anderson
Scott
S
Yates
John
J
Washburn
Michael P
MP
Workman
Jerry L
JL
D000107:Acetylation; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000502:physiology; D011401:Promoter Regions, Genetic; D021122:Protein Subunits / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
C098733:ASH1 protein, S cerevisiae; D004268:DNA-Binding Proteins; D009097:Multienzyme Complexes; D021122:Protein Subunits; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C089979:UME6 protein, S cerevisiae; D006655:Histone Deacetylases