title: PpNrg1 is a transcriptional repressor for glucose and glycerol repression of AOX1 promoter in methylotrophic yeast Pichia pastoris.
abstract: OBJECTIVE
  The regulator in glycerol repression of Pichia pastoris AOX1 promoter (P AOX1 ) is still unclear.
  
  
  RESULTS
  A Cys2His2 zinc finger transcriptional repressor PpNrg1 localized to nucleus and participated in the repression of P AOX1 in P. pastoris in glucose and glycerol. Quantitative real-time PCR revealed that PpNrg1 repressed expression of numerous genes involved in methanol utilization and peroxisome biogenesis in 0.02 % glucose and 1 % (v/v) glycerol. Electrophoretic mobility shift assay and DNase I footprinting assay revealed that PpNrg1 bound to five sites of P AOX1 , including two binding sites of PpMxr1, which is an indispensable activator of P AOX1 in P. pastoris.
  
  
  CONCLUSIONS
  Transcriptional repressor PpNrg1 suppresses P AOX1 in glucose and glycerol by directly binding to five sites of P AOX1 , including two binding sites of transcriptional activator PpMxr1.
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China. jxwangxl@126.com.
authors.forename: Xiaolong
authors.initials: X
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China. cmh022199@ecust.edu.cn.
authors.forename: Menghao
authors.initials: M
authors.lastname: Cai
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China. soshilei@qq.com.
authors.forename: Lei
authors.initials: L
authors.lastname: Shi
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China. wangqisam@163.com.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China. ywsy_bn@126.com.
authors.forename: Jinxiang
authors.initials: J
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China. lazeewang@gmail.com.
authors.forename: Jinjia
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China. mianzhou@ecust.edu.cn.
authors.forename: Mian
authors.initials: M
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China. xszhou@ecust.edu.cn.
authors.forename: Xiangshan
authors.initials: X
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China. yxzhang@ecust.edu.cn.|Shanghai Collaborative Innovation Center for Biomanufacturing (SCICB), Shanghai, 200237, China. yxzhang@ecust.edu.cn.
authors.forename: Yuanxing
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D018983:DNA Footprinting; D024202:Electrophoretic Mobility Shift Assay; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000378:metabolism*; D005990:Glycerol / Q000378:metabolism*; D010843:Pichia / Q000235:genetics* / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D060888:Real-Time Polymerase Chain Reaction; D012097:Repressor Proteins / Q000378:metabolism*
chemical_list: D012097:Repressor Proteins; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D005947:Glucose; D005990:Glycerol
keywords: AOX1 promoter; Catabolite repression; Glucose; Glycerol; Pichia pastoris; PpNrg1; Transcriptional regulation