ABSTRACT
We investigated inhibition of Moloney leukemia virus 10 (MOV10) upon xenotropic murine leukemia virus-related virus (XMRV) and made a preliminary study of the mechanism of action. Using transfection, infection, western blotting and real-time polymerase chain reaction, we found that MOV10 inhibited XMRV replication. Using MOV10 overexpressed in viral producer cells, MOV10 was shown to reduce the infectivity of XMRV. MOV10 could be incorporated into XMRV, suggesting that MOV10 could undergo encapsidation by XMRV during viral assembly. MOV10 could also restrict the DNA production of XMRV in target cells. We found that the putative RNA-helicase domain of MOV10 maintained most of its XMRV inhibition. These results suggest that MOV10 could be required during the retroviral lifecycle. Perturbation of MOV10 disrupts the generation of infectious viral particles, suggesting that MOV10 has broad antiretroviral activity. Hence, MOV10 could be actively involved in host defense against retroviral infection.
Subject(s)
Humans , Moloney murine leukemia virus , Physiology , RNA Helicases , Physiology , Virus ReplicationABSTRACT
BST-2 plays an important role in host innate immune response via inhibiting the release of HIV-1. HIV-1 accessory protein Vpu can interact with BST-2 through its transmembrane domains, degrade BST-2, and decrease BST-2 that are transported to the cell surface, thus anti-virus function of BST-2 is antagonized. In our study, we constructed plasmid RB connecting Rluc to the N-termimal of BST-2, and plasmid VE connecting EYFP to the C-terminal of Vpu. The two fusion proteins were co-expressed in 293 cells, and the interaction between the two proteins was detected via BRET method. And we further established a stable 293 cell line of dual-expression. By using BRET method, and the interaction between BST-2 and Vpu transmembrane domain as the target, a high-throughput screening assay was created that was expected to seek novel interaction inhibitors.
Subject(s)
Humans , Antigens, CD , Chemistry , Genetics , Metabolism , Cell Line , GPI-Linked Proteins , Chemistry , Genetics , Metabolism , HIV Infections , Genetics , Metabolism , Virology , HIV-1 , Genetics , Metabolism , High-Throughput Screening Assays , Methods , Human Immunodeficiency Virus Proteins , Genetics , Metabolism , Protein Binding , Protein Structure, Tertiary , Viral Regulatory and Accessory Proteins , Genetics , MetabolismABSTRACT
Recently, BST-2 has been identified as an effective cellular factor that prevents the release of human immunodeficiency virus type 1 and other enveloped viruses by tethering virus particles to the cell surface. Here, we showed that the production of HIV-1 virus-like particles was markedly inhibited by BST-2. Both the transient and stable expressing of BST-2 had the same function and Vpu rescued the release of HIV-1 VLP in the presence of human BST-2. Consistent with a direct tethering mechanism, we confirmed that proteolysis releases restricted virions and further showed that this removed the ectodomain of BST-2 from the cell surface.