文献类型：期刊文献

英文题名：Kinase Screening in Pichia pastoris Identified Promising Targets Involved in Cell Growth and Alcohol Oxidase 1 Promoter (PAOX1) Regulation

作者：Shen, Wei[1];Kong, Chuixing[1];Xue, Ying[1];Liu, Yiqi[1];Cai, Menghao[1];Zhang, Yuanxing[1,2];Jiang, Tianyi[3];Zhou, Xiangshan[1];Zhou, Mian[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai, Peoples R China;[2]SCICB, Shanghai, Peoples R China;[3]Roche R&D Ctr China Ltd, Shanghai, Peoples R China

年份：2016

卷号：11

期号：12

外文期刊名：PLOS ONE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000389587100180)】；

基金：This work was sponsored by Chinese National High Technology Research and Development Program (2014AA093501), National Natural Science Foundation of China (31600056), National Special Fund for State Key Laboratory of Bioreactor (2060204), and Shanghai Pujiang Program (15PJ1401600). Roche R&D (China) Ltd provided support in the form of salary for author [TJ], but did not have additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of the author are articulated in the "author contributions" section.

语种：英文

摘要：As one of the most commonly used eukaryotic recombinant protein expression systems, P. pastoris relies heavily on the AOX1 promoter (P-AOX1), which is strongly induced by methanol but strictly repressed by glycerol and glucose. However, the complicated signaling pathways involved in P-AOX1 regulation when supplemented with different carbon sources are poorly understood. Here we constructed a kinase deletion library in P. pastoris and identified 27 mutants which showed peculiar phenotypes in cell growth or PAOX1 regulation. We analyzed both annotations and possible functions of these 27 targets, and then focused on the MAP kinase Hog1. In order to locate its potential downstream components, we performed the phosphoproteome analysis on glycerol cultured WT and Delta hog1 strains and identified 157 differentially phosphorylated proteins. Our results identified important kinases involved in P. pastoris cell growth and P-AOX1 regulation, which could serve as valuable targets for further mechanistic studies.