regulating repression roles for the sir4 n-terminus in linker dna protection and stabilization of epigenetic states调节压抑的角色sir4 n端连接器dna保护和稳定的表观遗传状态.pdfVIP

Regulating Repression: Roles for the Sir4 N-Terminus in Linker DNA Protection and Stabilization of Epigenetic States 1 1 1,2 1 1¤ Stephanie Kueng , Monika Tsai-Pflugfelder , Mariano Oppikofer , Helder C. Ferreira , Emma Roberts , 1 1 1 1,2 Chinyen Tsai , Tim-Christoph Roloff , Ragna Sack , Susan M. Gasser * 1 Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland, 2 Faculty of Natural Sciences, University of Basel, Basel, Switzerland Abstract Silent information regulator proteins Sir2, Sir3, and Sir4 form a heterotrimeric complex that represses transcription at subtelomeric regions and homothallic mating type (HM) loci in budding yeast. We have performed a detailed biochemical and genetic analysis of the largest Sir protein, Sir4. The N-terminal half of Sir4 is dispensable for SIR–mediated repression of HM loci in vivo, except in strains that lack Yku70 or have weak silencer elements. For HM silencing in these cells, the C- terminal domain (Sir4C, residues 747–1,358) must be complemented with an N-terminal domain (Sir4N; residues 1–270), expressed either independently or as a fusion with Sir4C. Nonetheless, recombinant Sir4C can form a complex with Sir2 and Sir3 in vitro, is catalytically active, and has sedimentation properties similar to a full-length Sir4-containing SIR complex. Sir4C-containing SIR complexes bind nucleosomal arrays and protect linker DNA from nucleolytic digestion, but less effectively than wild-type SIR complexes. Consist