HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 VIF(-) MUTANT PARTICLES FROM RESTRICTIVE CELLS - ROLE OF VIF IN CORRECT PARTICLE ASSEMBLY AND INFECTIVITY

Disruption of the vif gene of human immunodeficiency virus (HIV) type 1 affects virus infectivity to various degrees, depending on the T-cell line used. We have concentrated our studies on true phenotypic Vif(-) mutant particles produced from CEMx174 or H9 cells. In a single round of infection, Vif(-) virus is approximately 25 (from CEMx174 cells) to 100 (from H9 cells) times less infectious than wild-type virus produced from these cells or than the Vif(-) mutant produced from HeLa cells. Vif(-) virions recovered from restrictive cells, but not from permissive cells, are abnormal both in terms of morphology and viral protein content. Notably, they contain much reduced quantities of envelope proteins and altered quantities of Gag and Pol proteins. Although wild-type and Vif(-) virions from restrictive cells contain similar quantities of viral RNA, no viral DNA synthesis was detectable after acute infection of target cells with phenotypically Vif(-) virions. To examine the possible role of Vif(-) in viral entry, attempts were made to rescue the Vif(-) defect in H9 cells by pseudotyping Vif(+) and Vif(-) HIV particles with amphotropic murine leukemia virus envelope. Vif(-) particles produced in the presence of HIV envelope could not be propagated when pseudotyped. In contrast, when only the murine leukemia virus envelope was present, significant propagation of Vif(-) HIV particles could be detected. These results demonstrate that Vif is required for proper assembly of the viral particle and for efficient HIV Env-mediated infection of target cells.