Profiling human protein degradome delineates cellular responses to proteasomal inhibition and reveals a feedback mechanism in regulating proteasome homeostasis  

作　　者：Tao Yu Yonghui Tao Meiqiang Yang Peng Chen Xiaobo Gao Yanbo Zhang Tao Zhang Zi Chen Jian Hou Yan Zhang Kangcheng Ruan Hongyan Wang Ronggui Hu 

机构地区：[1]State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China [2]Graduate School, Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, Chin [3]State Key Laboratory of Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China [4]Department of Laboratory Medicine, Hua-Shan Hospital, Fudan University, 12 Road Wulumuqi middle Road, Shanghai 200040, China [5]Department of Hematology, Hua-Shan Hospital, Fudan University, 12 Road Wulumuqi middle Road, Shanghai 200040, China [6]Department of Hematology, Changzheng Hospital, The Second Military Medical University, 415 Fengyang Road, Shanghai 200003, China [7]Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China [8]Cancer Research Center, SIBS-Xuhui Central Hospital, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China

出　　处：《Cell Research》2014年第10期1214-1230,共17页细胞研究（英文版）

基　　金：We are grateful to Dr W Bian and her team in Cell Biology Core facility of SIBCB for their excellent support in flow cytometry analysis and cell sorting, and Drs L Lin, Y Xia and H Xiao of National Center for Microarrays for their help in microarray analysis. We are particularly grateful to Dr Hsueh-Chi Yen at Institute of Mol Biol, Academia Sinica, and Dr Ivan Dikic of Goethe University for advice, and Drs Dangsheng Li, Lijian Hui and Jiarui Wu of SIBCB, and Dr Shirley Diamond of California Institute of Tech- nology for helpful discussion. We also thank Dr Ya-lan Wu and all other members of the Hu lab for support. This work was supported by the National Natural Science Foundation of China （31270828, 31070678, 81170491, 81072070 and 81470360）, the 100 Talents award from CAS to RH and the Ministry of Science and Technology, China （2010CB912100, 2012CB910800 and 2013CB910900 to RH, and 2011CB915501 to KR）. RH was also supported by a Sanofi-aventis SIBS Young Investigator award and funding from the Cancer Center of Xuhui Central Hospital （CCR2012003）, Shanghai Institute of Neurosciences （SKLN-201206） and the Instrument Developing Project of the Chinese Academy of Sciences （YZ201339）.

摘　　要：Global change in protein turnover （protein degradome） constitutes a central part of cellular responses to intrin- sic or extrinsic stimuli. However, profiling protein degradome remains technically challenging. Recently, inhibition of the proteasome, e.g., by using bortezomib （BTZ）, has emerged as a major chemotherapeutic strategy for treating multiple myeloma and other human malignancies, but systematic understanding of the mechanisms for BTZ drug action and tumor drug resistance is yet to be achieved. Here we developed and applied a dual-fluorescence-based Protein Turnover Assay （ProTA） to quantitatively profile global changes in human protein degradome upon BTZ-indueed proteasomal inhibition. ProTA and subsequent network analyses delineate potential molecular basis for BTZ action and tumor drug resistance in BTZ chemotherapy. Finally, combined use of BTZ with drugs targeting the ProTA-identified key genes or pathways in BTZ action reduced BTZ resistance in multiple myeloma cells. Remarkably, BTZ stabilizes proteasome subunit PSMCI and proteasome assembly factor PSMD10, suggesting a previously under-appreciated mechanism for regulating proteasome homeostasis. Therefore, ProTA is a novel tool for profiling human protein degradome to elucidate potential mechanisms of drug action and resistance, which might facilitate therapeutic development targeting proteostasis to treat human disorders.

关 键 词：BORTEZOMIB combination therapy PROTEOSTASIS protein degradome proteasome homeostasis N-end role ubiquitin 

分 类 号：Q55[生物学—生物化学] TS201.21[轻工技术与工程—食品科学]