LEDGF/p75 determines cellular trafficking of diverse lentiviral but not murine oncoretroviral integrase proteins and is a component of functional lentiviral preintegration complexes.
Human immunodeficiency virus type 1 (HIV-1), feline immunodeficiency virus (FIV), and Moloney murine leukemia virus (MoMLV) integrases were stably expressed to determine their intracellular trafficking. Each lentiviral integrase localized to cell nuclei in close association with chromatin while the murine oncoretroviral integrase was cytoplasmic. Fusions of pyruvate kinase to the lentiviral integrases did not reveal transferable nuclear localization signals. The intracellular trafficking of each was determined instead by the transcriptional coactivator LEDGF/p75, which was required for nuclear localization. Stable small interfering RNA expression eliminated detectable LEDGF/p75 expression and caused dramatic, stable redistribution of each lentiviral integrase from nucleus to cytoplasm while the distribution of MoMLV integrase was unaffected. In addition, endogenous LEDGF/p75 coimmunoprecipitated specifically with each lentiviral integrase. In vitro integration assays with preintegration complexes (PICs) showed that endogenous LEDGF/p75 is a component of functional HIV-1 and FIV PICs. However, HIV-1 and FIV infection and replication in LEDGF/p75-deficient cells was equivalent to that in control cells, whether cells were dividing or growth arrested. Two-long terminal repeat circle accumulation in nondividing cell nuclei was also equivalent to that of LEDGF/p75 wild-type cells. Virions produced in LEDGF/p75-deficient cells had normal infectivity. We conclude that LEDGF/p75 fully accounts for cellular trafficking of diverse lentiviral, but not oncoretroviral, integrases and is the main lentiviral integrase-to-chromatin tethering factor. While lentiviral PIC nuclear import is unaffected by LEDGF/p75 knockdown, this protein is a component of functional lentiviral PICs. A role in HIV-1 integration site distribution merits investigation.
Llano
Manuel
M
Molecular Medicine Program, Mayo Clinic College of Medicine, Rochester, Minnesota 55905, USA.
Vanegas
Maria
M
Fregoso
Oliver
O
Saenz
Dyana
D
Chung
Susan
S
Peretz
Mary
M
Poeschla
Eric M
EM
D021581:Active Transport, Cell Nucleus; D002460:Cell Line; D002843:Chromatin / Q000378:metabolism; D015870:Gene Expression; D015694:Gene Products, rev / Q000502:physiology; D015497:HIV-1 / Q000737:chemistry / Q000201:enzymology / Q000502:physiology; D006801:Humans; D016089:Immunodeficiency Virus, Feline / Q000737:chemistry / Q000201:enzymology / Q000502:physiology; D019426:Integrases / Q000235:genetics / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D007438:Introns / Q000235:genetics; D016086:Lentivirus / Q000737:chemistry* / Q000201:enzymology* / Q000502:physiology; D019913:Nuclear Localization Signals; D009858:Oncogenic Viruses / Q000201:enzymology*; D011485:Protein Binding; D012190:Retroviridae / Q000201:enzymology*; D016662:Virus Integration / Q000502:physiology*; D014779:Virus Replication / Q000502:physiology; D054321:rev Gene Products, Human Immunodeficiency Virus
D002843:Chromatin; D015694:Gene Products, rev; D036341:Intercellular Signaling Peptides and Proteins; D019913:Nuclear Localization Signals; C119380:lens epithelium-derived growth factor; D054321:rev Gene Products, Human Immunodeficiency Virus; D019426:Integrases