摘要
Human immunodeficiency virus-1 （HIV-1）encodes 15 viral proteins. Protein-protein interactions play a large role in the function of these proteins. In this study, we attempted to identify novel interactions between the HIV-1 proteins to better understand the role played by viral protein-protein interactions in the life cycle of HIV-I. Genes encoding the 15 viral proteins from the HIV-1 strain AD8 were inserted into the plasmids of a yeast two-hybrid system. By screening 120 pairs of proteins, interactions between seven pairs were found. This led to the discovery of an interaction between the HIV-1 proteins integrase （IN） and glycoprotein 41 （gp41）, which was confirmed by both co-immunoprecipitation （Co-IP） assays and fluorescence resonance energy transfer （FRET） imaging in live cells. In addition, it was found that the amino acids at positions 76-100 of gp41 are required for it to bind to IN. Deletion of this region from gp41 prevented its interaction with IN and reduced the production of HIV-1 in 293T cells. This study provides new information on HIV-1 protein-protein interactions which improves the understanding of the biological functions of gp41 and IN during the virus life cycle.
Human immunodeficiency virus-1(HIV-1) encodes 15 viral proteins. Protein-protein interactions play a large role in the function of these proteins. In this study, we attempted to identify novel interactions between the HIV-1 proteins to better understand the role played by viral protein-protein interactions in the life cycle of HIV-1. Genes encoding the 15 viral proteins from the HIV-1 strain AD8 were inserted into the plasmids of a yeast two-hybrid system. By screening 120 pairs of proteins, interactions between seven pairs were found. This led to the discovery of an interaction between the HIV-1 proteins integrase(IN) and glycoprotein 41(gp41), which was confirmed by both co-immunoprecipitation(Co-IP) assays and fluorescence resonance energy transfer(FRET)imaging in live cells. In addition, it was found that the amino acids at positions 76–100 of gp41 are required for it to bind to IN. Deletion of this region from gp41 prevented its interaction with IN and reduced the production of HIV-1 in 293 T cells. This study provides new information on HIV-1protein-protein interactions which improves the understanding of the biological functions of gp41 and IN during the virus life cycle.
基金
supported by the National Nano Project(grant no.2011CB933600)
the National Natural Science Foundation of China(grant no.31470269)
supported by the Institute of Biophysics and the National Laboratory of Biomacromolecules at the Chinese Academy of Sciences(CAS)
