Integration is required for productive infection of monocyte-derived macrophages by human immunodeficiency virus type 1.
Certain human immunodeficiency virus type 1 (HIV-1) isolates are able to productively infect nondividing cells of the monocyte/macrophage lineage. We have used a molecular genetic approach to construct two different HIV-1 integrase mutants that were studied in the context of an infectious, macrophage-tropic HIV-1 molecular clone. One mutant, HIV-1 delta D(35)E, containing a 37-residue deletion within the central, catalytic domain of integrase, was noninfectious in both peripheral blood mononuclear cells and monocyte-derived macrophages. The HIV-1 delta D(35)E mutant, however, exhibited defects in the assembly and/or release of progeny virions in transient transfection assays, as well as defects in entry and/or viral DNA synthesis during the early stages of monocyte-derived macrophage infection. The second mutant, HIV-1D116N/8, containing a single Asp-to-Asn substitution at the invariant Asp-116 residue of integrase, was also noninfectious in both peripheral blood mononuclear cells and monocyte-derived macrophages but, in contrast to HIV-1 delta D(35)E, was indistinguishable from wild-type virus in reverse transcriptase production. PCR analysis indicated that HIV-1D116N/8 entered monocyte-derived macrophages efficiently and reverse transcribed its RNA but was unable to complete its replication cycle because of a presumed block to integration. These data are consistent with the hypothesis that integration is an obligate step in productive HIV-1 infection of activated peripheral blood mononuclear cells and primary human macrophage cultures.
Englund
G
G
Laboratory of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
Theodore
T S
TS
Freed
E O
EO
Engelman
A
A
Martin
M A
MA
D000595:Amino Acid Sequence; D001483:Base Sequence; D002478:Cells, Cultured; D004254:DNA Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D015751:Genes, pol; D015497:HIV-1 / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D019426:Integrases; D008264:Macrophages / Q000821:virology*; D008969:Molecular Sequence Data; D009000:Monocytes / Q000821:virology; D017384:Sequence Deletion; D016662:Virus Integration / Q000235:genetics / Q000502:physiology*; D014779:Virus Replication / Q000235:genetics / Q000502:physiology
D017931:DNA Primers; D004279:DNA, Viral; D004254:DNA Nucleotidyltransferases; D019426:Integrases