title: Transportin 3 and importin α are required for effective nuclear import of HIV-1 integrase in virus-infected cells.
abstract: Unlike other retroviruses, human immunodeficiency virus type-1 (HIV-1) can infect terminally differentiated cells, due to the ability of its pre-integration complex (PIC) to translocate via the host nuclear pore complex (NPC). The PIC Nuclear import has been suggested to be mediated by the viral integrase protein (IN), via either the importin α or transportin 3 (TNPO3/transportin-SR2) pathways.We show that in virus-infected cells, IN interacts with both importin α and TNPO3, simultaneously or separately, suggesting a multiple use of nuclear import pathways. Disruption of either the IN-importin α or IN-TNPO3 complexes in virus-infected cells by specific cell-permeable-peptides resulted in inhibition of IN and viral cDNA nuclear import. Here we show that peptides which disrupt either one of these complexes block virus infection, indicating involvement of both pathways in efficient viral replication. Formation of IN-importin α and IN-TNPO3 complexes has also been observed in IN-transfected cultured cells. Using specific peptides, we demonstrate that in transfected cells but not in virus infected cells the importin α pathway overrides that of TNPO3. The IN-importin α and IN-TNPO3 complexes were not observed in virus-infected Rev-expressing cells, indicating the Rev protein's ability to disrupt both complexes.Our work suggests that IN nuclear import requires the involvement of both importin α and TNPO3. The ability to inhibit nuclear import of the IN-DNA complex and consequently, virus infection by peptides that interrupt IN's interaction with either importin α or TNPO3 indicates that for efficient infection, nuclear import of IN should be mediated by both nuclear-import receptors.
authors.affiliation: Department of Biological Chemistry, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Safra Campus, Givat Ram, Jerusalem 91904, Israel.
authors.forename: Aviad
authors.initials: A
authors.lastname: Levin
authors.forename: Zvi
authors.initials: Z
authors.lastname: Hayouka
authors.forename: Assaf
authors.initials: A
authors.lastname: Friedler
authors.forename: Abraham
authors.initials: A
authors.lastname: Loyter
mesh_terms: D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D000998:Antiviral Agents / Q000138:chemical synthesis / Q000494:pharmacology; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D004279:DNA, Viral / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D019427:HIV Integrase / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D008969:Molecular Sequence Data; D022022:Nuclear Pore / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology; D011485:Protein Binding; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects; D028901:alpha Karyopherins / Q000235:genetics / Q000378:metabolism*; D028961:beta Karyopherins / Q000235:genetics / Q000378:metabolism*; D054321:rev Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism
chemical_list: D000998:Antiviral Agents; D018076:DNA, Complementary; D004279:DNA, Viral; D010455:Peptides; C546476:TNPO3 protein, human; D028901:alpha Karyopherins; D028961:beta Karyopherins; D054321:rev Gene Products, Human Immunodeficiency Virus; C514860:rev protein, Human Immunodeficiency Virus-1; D019427:HIV Integrase; C514801:p31 integrase protein, Human immunodeficiency virus 1