文献类型：期刊文献

英文题名：Mit1 Transcription Factor Mediates Methanol Signaling and Regulates the Alcohol Oxidase 1 (AOX1) Promoter in Pichia pastoris

作者：Wang, Xiaolong[1];Wang, Qi[1];Wang, Jinjia[1];Bai, Peng[1];Shi, Lei[1];Shen, Wei[1];Zhou, Mian[1];Zhou, Xiangshan[1];Zhang, Yuanxing[1,2];Cai, Menghao[1]

机构：[1]E China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]SCICB, Shanghai 200237, Peoples R China

年份：2016

卷号：291

期号：12

起止页码：6245

外文期刊名：JOURNAL OF BIOLOGICAL CHEMISTRY

收录：;EI(收录号：20161902345053);WOS:【SCI-EXPANDED(收录号:WOS:000372894200016)】；

基金：This work was supported by Grant 2060204 from the National Special Fund for State Key Laboratory of Bioreactor Engineering and Grant 2014AA093501 from the Chinese National High Technology Research and Development Program. The authors declare that they have no conflicts of interest with the contents of this article.

语种：英文

外文关键词：DNA-protein interaction; protein domain; signal transduction; transcription coactivator; transcription regulation; AOX1 promoter; Mit1; Pichia pastoris; glycerol repression; methanol induction

摘要：The alcohol oxidase 1 (AOX1) promoter (P-AOX1) of Pichia pastoris is the most powerful and commonly used promoter for driving protein expression. However, mechanisms regulating its transcriptional activity are unclear. Here, we identified a Zn(II)(2)Cys(6)-type methanol-induced transcription factor 1 (Mit1) and elucidated its roles in regulating P-AOX1 activity in response to glycerol and methanol. Mit1 regulated the expression of many genes involved in methanol utilization pathway, including AOX1, but did not participate in peroxisome proliferation and transportation of peroxisomal proteins during methanol metabolism. Structural analysis of Mit1 by performing domain deletions confirmed its specific and critical role in the strict repression of P-AOX1 in glycerol medium. Importantly, Mit1, Mxr1, and Prm1, which positively regulated P-AOX1 in response to methanol, were bound to P-AOX1 at different sites and did not interact with each other. However, these factors cooperatively activated P-AOX1 through a cascade. Mxr1 mainly functioned during carbon derepression, whereas Mit1 and Prm1 functioned during methanol induction, with Prm1 transmitting methanol signal to Mit1 by binding to the MIT1 promoter (P-MIT1), thus increasingly expressing Mit1 and subsequently activating P-AOX1.