文献类型：期刊文献

英文题名：Transcriptome analysis of Δmig1Δmig2 mutant reveals their roles in methanol catabolism, peroxisome biogenesis and autophagy in methylotrophic yeast Pichia pastoris

作者：Shi, Lei[1];Wang, Xiaolong[1];Wang, Jinjia[1];Zhang, Ping[1];Qi, Fei[1];Cai, Menghao[1];Zhang, Yuanxing[1,2];Zhou, Xiangshan[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2018

卷号：40

期号：4

起止页码：399

外文期刊名：GENES & GENOMICS

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000429036600007)】；

基金：This work was supported by Shanghai Science and Technology Innovation Plan (17JC1402400); Fundamental Research Funds for the Central Universities (22A201514040) and Grants of Young and Middle-aged Leading Science and Technology Innovation Talents from Ministry of Science and Technology of China. The authors are grateful to Prof. James M. Cregg, BioGrammatics, Inc. and Keck Graduate Institute of Applied Life Science, for helpful suggestions. The authors also thank Shanghai Huaguan Biochip Co. Ltd. for the help of RNA-seq data analysis.

语种：英文

外文关键词：Pichia pastoris; RNA-seq; Mig; Alcohol oxidase; Catabolite repression

摘要：Two catabolite repressor genes (MIG1 and MIG2) were previously identified in Pichia pastoris, and the derepression of alcohol oxidase (AOX) expression was realized in Delta mig1 or Delta mig1 Delta mig2 mutants grown in glycerol, but not in glucose. In this study, genome-wide RNA-seq analysis of Delta mig1 Delta mig2 and the wild-type strain grown in glycerol revealed that the expression of numerous genes was greatly altered. Nearly 7% (357 genes) of approximately 5276 genes annotated in P. pastoris were significantly upregulated, with at least a two-fold differential expression in Delta mig1 Delta mig2; the genes were mainly related to cell metabolism. Approximately 23% (1197 genes) were significantly downregulated; these were mainly correlated with the physiological characteristics of the cell. The methanol catabolism and peroxisome biogenesis pathways were remarkably enhanced, and the genes AOX1 and AOX2 were upregulated higher than 30-fold, which was consistent with the experimental results of AOX expression. The Mig proteins had a slight effect on autophagy when cells were grown in glycerol. The expression analysis of transcription factors showed that deletion of MIG1 and MIG2 significantly upregulated the binding of an essential transcription activator, Mit1p, with the AOX1 promoter, which suggested that Mig proteins might regulate the AOX1 promoter through the regulation of Mit1p. This work provides a reference for the further exploration of the methanol induction and catabolite repression mechanisms of AOX expression in methylotrophic yeasts.