Wang Xiaolong, Cai Menghao, Shi Lei, Wang Qi, Zhu Jinxiang, Wang Jinjia, Zhou Mian, Zhou Xiangshan, Zhang Yuanxing
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China.
Shanghai Collaborative Innovation Center for Biomanufacturing (SCICB), Shanghai, 200237, China.
Biotechnol Lett. 2016 Feb;38(2):291-8. doi: 10.1007/s10529-015-1972-4. Epub 2015 Oct 13.
The regulator in glycerol repression of Pichia pastoris AOX1 promoter (P AOX1 ) is still unclear.
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.
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.
毕赤酵母醇氧化酶1启动子（PAOX1）甘油抑制中的调节因子仍不清楚。
一种Cys2His2锌指转录抑制因子PpNrg1定位于细胞核，并参与毕赤酵母在葡萄糖和甘油条件下对PAOX1的抑制。定量实时PCR显示，PpNrg1在0.02%葡萄糖和1%（v/v）甘油条件下抑制了许多参与甲醇利用和过氧化物酶体生物发生的基因的表达。电泳迁移率变动分析和DNase I足迹分析显示，PpNrg1与PAOX1的五个位点结合，其中包括PpMxr1的两个结合位点，PpMxr1是毕赤酵母中PAOX1不可或缺的激活因子。
转录抑制因子PpNrg1通过直接结合PAOX1的五个位点（包括转录激活因子PpMxr1的两个结合位点）在葡萄糖和甘油条件下抑制PAOX1。
