Zhao Rongmin, Davey Mike, Hsu Ya-Chieh, Kaplanek Pia, Tong Amy, Parsons Ainslie B, Krogan Nevan, Cagney Gerard, Mai Duy, Greenblatt Jack, Boone Charles, Emili Andrew, Houry Walid A
Department of Biochemistry, Medical Sciences Building, 1 King's College Circle, University of Toronto, Toronto, ON, M5S 1A8, Canada.
Cell. 2005 Mar 11;120(5):715-27. doi: 10.1016/j.cell.2004.12.024.
Physical, genetic, and chemical-genetic interactions centered on the conserved chaperone Hsp90 were mapped at high resolution in yeast using systematic proteomic and genomic methods. Physical interactions were identified using genome-wide two hybrid screens combined with large-scale affinity purification of Hsp90-containing protein complexes. Genetic interactions were uncovered using synthetic genetic array technology and by a microarray-based chemical-genetic screen of a set of about 4700 viable yeast gene deletion mutants for hypersensitivity to the Hsp90 inhibitor geldanamycin. An extended network, consisting of 198 putative physical interactions and 451 putative genetic and chemical-genetic interactions, was found to connect Hsp90 to cofactors and substrates involved in a wide range of cellular functions. Two novel Hsp90 cofactors, Tah1 (YCR060W) and Pih1 (YHR034C), were also identified. These cofactors interact physically and functionally with the conserved AAA(+)-type DNA helicases Rvb1/Rvb2, which are key components of several chromatin remodeling factors, thereby linking Hsp90 to epigenetic gene regulation.
利用系统的蛋白质组学和基因组学方法，在酵母中以高分辨率绘制了以保守伴侣蛋白Hsp90为中心的物理、遗传和化学遗传相互作用图谱。通过全基因组双杂交筛选结合含Hsp90蛋白复合物的大规模亲和纯化来鉴定物理相互作用。利用合成遗传阵列技术以及基于微阵列的化学遗传筛选方法，对一组约4700个存活的酵母基因缺失突变体进行筛选，以检测其对Hsp90抑制剂格尔德霉素的超敏感性，从而发现遗传相互作用。发现一个由198个推定的物理相互作用以及451个推定的遗传和化学遗传相互作用组成的扩展网络，将Hsp90与参与广泛细胞功能的辅助因子和底物联系起来。还鉴定出两个新的Hsp90辅助因子，即Tah1（YCR060W）和Pih1（YHR034C）。这些辅助因子与保守的AAA(+)型DNA解旋酶Rvb1/Rvb2在物理和功能上相互作用，而Rvb1/Rvb2是几种染色质重塑因子的关键组分，从而将Hsp90与表观遗传基因调控联系起来。
