Study on the interaction between hepatitis virus C nonstructural protein 4A and calcium modulating cyclophipin ligand by in vivo coimmunoprecipitation
- VernacularTitle:丙型肝炎病毒非结构蛋白4A与钙离子信号调节亲环素配体的相互作用
- Author:
Yong-Qian CHENG
;
Jun CHENG
;
Lin WANG
;
Yan LIU
;
Dong-Ping XU
;
Yan-Wei ZHONG
;
Jian-Hui QU
;
Jiu-Zeng DAI
;
Xiao-Dong LI
;
- Publication Type:Journal Article
- Keywords:
Hepacivirus;
Viral nonstructural proteins;
Calcium signaling;
Signal transduc-tion;
Cyclophilins ligand;
Yeasts;
Hybridization;
Immunoprecipitation
- From:
Chinese Journal of Infectious Diseases
2000;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To prove the interaction between hepatitis virus C(HCV)nonstruetural protein 4A(HCV NS4A)and calcium modulating cyclophilin tigand(CAML)with yeast-two hybrid- ization and coimmunoprecipitation.Methods The gene encoding CAML was cloned,and subcloned into the yeast expression vector pGADT7 and eucell expression vector pcDNA3.1/His-A.The back- cross test between HCV NS4A and CAML was performed in yeast cells.After that,the pCMV-Myc/ NS4A plasmid and pcDNA3.1/His-A-CAML plasmid were co transfected into 293 cells and,then, coimmunoprecipitation and Western blot were performed.Results The gene encoding CAML was cloned sucessfully,and then the gene was subcloned into yeast expression vectors,pGADT7.After the interaction between NS4A and CAML was ensured in yeast cells,the eukaryotic expression vec- tors of NS4A and CAML were constructed and their interaction was ensured again by Co-immunopre- cipitation.Conclusions The interaction between HCV NS4A and CAML is proved.CAML is one of the proteins involved in Ca~(2+)signaling,which suggests that the interaction of HCV NS4A and CAML may be a new clue of the chronic mechanism of HCV infection.Future studies will be required to de- fine the physiologic significance of this interaction.
