Mit1 Transcription Factor Mediates Methanol Signaling and Regulates the Alcohol Oxidase 1 (AOX1) Promoter in Pichia pastoris.

The Journal of Biological Chemistry
Xiaolong WangMenghao Cai

Abstract

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)2Cys6-type methanol-induced transcription factor 1 (Mit1) and elucidated its roles in regulating PAOX1 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 express...Continue Reading

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Citations

Oct 23, 2016·Microbial Cell Factories·Wei ShenMian Zhou
Jul 14, 2018·Biotechnology Letters·Jinxiang ZhuYuanxing Zhang
Jan 28, 2021·Animals : an Open Access Journal From MDPI·Jingyi JiangBichun Li
Apr 2, 2020·ACS Synthetic Biology·Jiao WenMenghao Cai
Jul 22, 2021·Biotechnology for Biofuels·Nai-Xin LinXiao-Wei Yu
Sep 1, 2021·Journal of Bioscience and Bioengineering·Takehiko Yoko-OYasunori Chiba

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Datasets Mentioned

BETA
CAY70887
AAO72735

Methods Mentioned

BETA
PCR
footprinting
Immunoprecipitation
ChIP
Pulldown
Two-hybrid
Fluorescence
protein assay
reverse transcription-PCR
RNA-Seq

Software Mentioned

- Pro Analyzer
Gel
ClustalX
Leica
BLAST
GeneDoc

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