2.表观遗传与疾病的课件.pptVIP

* * * * * The Strategy Purify Ajuba-associated protein complexes by large scale affinity chromatography combined with Mass spectrometry (MS) : stably express Flag-epitope tagged Ajuba in 293 cells, and the Ajuba associated protein complexes were immunoprecipitated with anti-Flag M2 beads. The components of the complexes were identified by MS. Affinity purification of Ajuba-interacting proteins from 293 cells Prmt5 is a type II Protein Arginine methyltransferase Type I: Prmt1, 2, 3, 4, and 6 Type II: Prmt5 E-box Zn Zn Zn Zn Zn Zn Zn Zn Zn SNAG Ajuba LIM Snail preLIM PRMT5 MEP50 E-cadherin Snail target genes Conclusion: Prmt5 is an effector recruited to Snail via an interaction with Ajuba Snail contains conserved 14-3-3 binding motifs 最近发现14-3-3能阅读组蛋白修饰密码 Snail selectively interacts with 14-3-3 isoforms Mutation of the 14-3-3 binding sites in Snail Mutation of the 14-3-3 binding motifs impairs the transcriptional repression activity of Snail The 14-3-3 binding sites in Snail are essential for induction of EMT in MCF10A cells Ajuba is a 14-3-3 interacting protein Summary Snail recruits Ajuba as a corepressor via its SNAG motif. Ajuba further recruits Prmt5 which confers the Snail complex the transcriptional repression activity by methylate histones 14-3-3 binds Ajuba and Snail, which is essential for the Snail complex to bind the target chromatin. PRC2 Suz12/Ezh2 H3K27me2/3 mSin3 HDAC1/2 Deacetylation LSD1 G9a Suv39H1 Demethylation H3K9me2 Ajuba Co-rest The carboxyl zinc fingers of Snail recruit ubiquitin E3 ligases Ring1A/B Cancer Research, 2014 The Zinc fingers of Snail bind to the RING domain of Ring1B Ring1A/B are essential for Snail-mediated repression and cell migration Snail enhances Ring1A/B binding to its target promoter PRC2 Suz12/Ezh2 H3K27me2/3 Ajuba Ezh2 is required for Snail/Ring1A/B to bind target promoter Ezh2 and Ring1A/B form two distinct complexes with Snail High levels of Snail , Ring1B and H2AK119Ub1 in PDAC H2AK119Ub1 is a biomarker predic