title: A clathrin, caveolae, and dynamin-independent endocytic pathway requiring free membrane cholesterol drives HIV-1 internalization and infection in polarized trophoblastic cells.
abstract: In human trophoblastic cells, a correlation between early endosomal trafficking of HIV-1 and virus infection was previously documented. However, if HIV-1 is massively internalized in these cells, the endocytic pathway(s) responsible for viral uptake is still undefined. Here we address this vital question. Amongst all the putative endocytic pathways present in polarized trophoblastic cells, we demonstrate that HIV-1 infection of these cells is independent of clathrin-mediated endocytosis and macropinocytosis. Importantly, treatment with the cholesterol-sequestering drug filipin severely impairs virus internalization, whereas the cholesterol-depleting compound methyl-beta-cyclodextrin has no impact on this pathway. Moreover, viral internalization is unaffected by overexpression of a mutant dynamin 2 or treatment with a kinase or tyrosine phosphatase inhibitor. Thus, HIV-1 infection in polarized trophoblastic cells occurs primarily via a clathrin, caveolae, and dynamin-independent pathway requiring free cholesterol. Notably, even though HIV-1 did not initially co-localize with transferrin, some virions migrate at later time points to transferrin-enriched endosomes, suggesting an unusual transit from the non-classical pathway to early endosomes. Finally, virus internalization in these cells does not involve the participation of microtubules but relies partly on actin filaments. Collectively these findings provide unprecedented information on the route of HIV-1 internalization in polarized human trophoblasts.
authors: Research Center in Infectious Diseases, CHUL Research Center, and Department of Medical Biology, Faculty of Medicine, Laval University, Quebec, Canada G1V 4G2.
authors: Gaël
authors: G
authors: Vidricaire
authors: Michel J
authors: MJ
authors: Tremblay
mesh_terms: D000199:Actins / Q000378:metabolism; D000328:Adult; D021941:Caveolae / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D016764:Cell Polarity; D002648:Child; D002784:Cholesterol / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D034281:Dynamins / Q000378:metabolism*; D004705:Endocytosis / Q000187:drug effects / Q000502:physiology*; D005260:Female; D005372:Filipin / Q000494:pharmacology; D015658:HIV Infections; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D021962:Membrane Microdomains / Q000737:chemistry / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism; D014327:Trophoblasts / Q000166:cytology / Q000378:metabolism / Q000821:virology; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D053586:Virus Internalization; D047392:beta-Cyclodextrins / Q000494:pharmacology
chemical_list: D000199:Actins; D002966:Clathrin; D014168:Transferrin; D014409:Tumor Necrosis Factor-alpha; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D005372:Filipin; D002784:Cholesterol; D034281:Dynamins