title: Kinase Screening in Pichia pastoris Identified Promising Targets Involved in Cell Growth and Alcohol Oxidase 1 Promoter (PAOX1) Regulation.
abstract: As one of the most commonly used eukaryotic recombinant protein expression systems, P. pastoris relies heavily on the AOX1 promoter (PAOX1), which is strongly induced by methanol but strictly repressed by glycerol and glucose. However, the complicated signaling pathways involved in PAOX1 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 Δhog1 strains and identified 157 differentially phosphorylated proteins. Our results identified important kinases involved in P. pastoris cell growth and PAOX1 regulation, which could serve as valuable targets for further mechanistic studies.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Shen
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
authors.forename: Chuixing
authors.initials: C
authors.lastname: Kong
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xue
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
authors.forename: Yiqi
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
authors.forename: Menghao
authors.initials: M
authors.lastname: Cai
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.|Shanghai Collaborative Innovation Center for Biomanufacturing (SCICB), Shanghai, China.
authors.forename: Yuanxing
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Roche R&D Center (China) Ltd, Pudong, Shanghai, China.
authors.forename: Tianyi
authors.initials: T
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
authors.forename: Xiangshan
authors.initials: X
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
authors.forename: Mian
authors.identifier: http://orcid.org/0000-0003-4226-6299
authors.initials: M
authors.lastname: Zhou
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D010843:Pichia / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development*; D011401:Promoter Regions, Genetic; D011494:Protein Kinases / Q000235:genetics*
chemical_list: D005656:Fungal Proteins; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D011494:Protein Kinases