Biochemical characterization and functional analysis of the POU transcription factor POU-M2 of Bombyx mori
POU-M2 is a homeodomain transcription factor which plays important roles in the development and silk synthesis of Bombyx mori.In this study,we expressed,purified and characterized POU-M2 and studied its transcription regulation on fibroin heavy chain gene of Bombyx mori.Gel filtration showed POU-M2 existed as a dimer in solution.Far-UV circular dichroism spectra indicated POU-M2 had a well-definedα-helix structure and theα-helix content was about 26.4%.The thermal unfolding transition of POU-M2 was a cooperative process.Tm,ΔH and ΔS were 45.15 ± 0.2℃,138.4 ± 0.5 KJ/mol and 0.4349 ± 0.04KJ/(mol· K),respectively.Western blotting analysis indicated the expression level of POU-M2 increased slightly from day 3 to day 7 of the fifth instar larvae in the posterior silk gland.POUM2 was positioned in the nucleus of cells.The luciferase reporter assay demonstrated POU-M2 could stimulate the promoter activity of fibroin heavy chain gene,and the activation effect was dependent on the amount of POU-M2.Our study suggested POU-M2 may be involved in the transcriptional regulation of fibroin heavy chain gene.These findings expand toward a better understanding of the structure of POU-M2 and its function in silk synthesis of Bombyx mori.
Bombyx mori POU-M2 biochemical characterization fibroin heavy chain transcriptional regulation
Liu lina Li yu Wang yejing Zhao peng Wei shuguang Li zhenzhen Chang huaipu He huawei
State Key Laboratory of Silkworm Genome Biology,Southwest University,Beibei,Chongqing 400715,China State Key Laboratory of Silkworm Genome Biology,Southwest University,Beibei,Chongqing 400715,China;K
国内会议
重庆
英文
173-183
2016-05-25（万方平台首次上网日期，不代表论文的发表时间）