The Cell Cycle Independence of HIV Infections Is Not Determined by Known Karyophilic Viral Elements

Abstract

Human immunodeficiency virus and other lentiviruses infect cells independent of cell cycle progression, but gammaretroviruses, such as the murine leukemia virus (MLV) require passage of cells through mitosis. This property is thought to be important for the ability of HIV to infect resting CD4+ T cells and terminally differentiated macrophages. Multiple and independent redundant nuclear localization signals encoded by HIV have been hypothesized to facilitate migration of viral genomes into the nucleus. The integrase (IN) protein of HIV is one of the HIV elements that targets to the nucleus; however, its role in nuclear entry of virus genomes has been difficult to describe because mutations in IN are pleiotropic. To investigate the importance of the HIV IN protein for infection of non-dividing cells, and to investigate whether or not IN was redundant with other viral signals for cell cycle-independent nuclear entry, we constructed an HIV-based chimeric virus in which the entire IN protein of HIV was replaced by that of MLV. This chimeric virus with a heterologous IN was infectious at a low level, and was able to integrate in an IN-dependent manner. Furthermore, this virus infected non-dividing cells as well as it infected dividing cells. Moreover, we used the chimeric HIV with MLV IN to further eliminate all of the other described nuclear localization signals from an HIV genome—matrix, IN, Viral Protein R, and the central polypurine tract—and show that no combination of the virally encoded NLS is essential for the ability of HIV to infect non-dividing cells. Human immunodeficiency virus can infect many cells irrespective of whether or not they are dividing, whereas some other retroviruses, such as the murine leukemia virus can only infect cells that are proliferating. This property is important for the ability of HIV to establish infections in critical cell types in infected people. Multiple and redundant signals encoded by HIV have been hypothesized to facilitate migration of viral genomes into the nucleus. However, here the authors eliminated all four described nuclear localizing signals from an HIV genome and show that no combination of these virally encoded signals is essential for the ability of HIV to infect non-dividing cells. They suggest that another step of the virus lifecycle, other than nuclear import, is the rate-limiting step that determines the cell cycle dependence/independence of retroviral infections.