title: Mit1 Transcription Factor Mediates Methanol Signaling and Regulates the Alcohol Oxidase 1 (AOX1) Promoter in Pichia pastoris.
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 expressing Mit1 and subsequently activating P AOX1.
authors.affiliation: From the State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China and.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Wang
authors.affiliation: From the State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China and.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: From the State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China and.
authors.forename: Jinjia
authors.initials: J
authors.lastname: Wang
authors.affiliation: From the State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China and.
authors.forename: Peng
authors.initials: P
authors.lastname: Bai
authors.affiliation: From the State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China and.
authors.forename: Lei
authors.initials: L
authors.lastname: Shi
authors.affiliation: From the State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China and.
authors.forename: Wei
authors.initials: W
authors.lastname: Shen
authors.affiliation: From the State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China and.
authors.forename: Mian
authors.initials: M
authors.lastname: Zhou
authors.affiliation: From the State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China and.
authors.forename: Xiangshan
authors.initials: X
authors.lastname: Zhou
authors.affiliation: From the State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China and the Shanghai Collaborative Innovation Center for Biomanufacturing (SCICB), Shanghai 200237, China.
authors.forename: Yuanxing
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: From the State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, China and cmh022199@ecust.edu.cn.
authors.forename: Menghao
authors.initials: M
authors.lastname: Cai
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001665:Binding Sites; D004794:Enzyme Repression; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D000432:Methanol / Q000378:metabolism*; D008969:Molecular Sequence Data; D020675:Peroxisomes / Q000378:metabolism; D010843:Pichia / Q000201:enzymology* / Q000235:genetics; D011401:Promoter Regions, Genetic; D021381:Protein Transport; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D014157:Transcription Factors / Q000502:physiology*
chemical_list: D005656:Fungal Proteins; D014157:Transcription Factors; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D000432:Methanol
keywords: AOX1 promoter; DNA-protein interaction; Mit1; Pichia pastoris; glycerol repression; methanol induction; protein domain; signal transduction; transcription coactivator; transcription regulation
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