The ubiquitin-vacuolar protein sorting system is selectively required during entry of influenza virus into host cells.
Influenza virus enters cells by endocytosis, and requires the low pH of the late endosome for successful infection. Here, we investigated the requirements for sorting into the multivesicular body pathway of endocytosis. We show that treatment of host cells with the proteasome inhibitors MG132 and lactacystin directly affects the early stages of virus replication. Unlike other viruses, such as retroviruses, influenza virus budding was not affected. The requirement for proteasome function was not shared by two other pH-dependent viruses: Semliki Forest virus and vesicular stomatitis virus. With MG132 treatment, incoming influenza viruses were retained in endosomes that partially colocalized with mannose 6-phosphate receptor, but not with classical markers of early or late endosomes. Colocalization was also observed with Rme-1, which is part of the recycling pathway of endocytosis. In addition, influenza virus entry was dependent on the vacuolar protein sorting pathway, as over-expression of dominant-negative hVPS4 caused arrest of viruses in endosome-like populations that partially colocalized with the hVPS4 protein. Overall, we conclude that influenza virus selectively requires the ubiquitin/vacuolar protein sorting pathway for entry into host cells, and that it must communicate with a specific cellular machinery for intracellular sorting during the initial phase of virus infection.
Khor
Rebecca
R
Department of Microbiology & Immunology, Cornell University, Ithaca NY 14853, USA.
McElroy
Lisa J
LJ
Whittaker
Gary R
GR
D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004705:Endocytosis; D056827:Endosomal Sorting Complexes Required for Transport; D011992:Endosomes / Q000378:metabolism; D005434:Flow Cytometry; D005799:Genes, Dominant; D049452:Green Fluorescent Proteins; D006388:Hemagglutinins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007976:Leupeptins / Q000494:pharmacology; D008164:Luminescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D008907:Mink; D009975:Orthomyxoviridae / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D021381:Protein Transport; D017527:Receptor, IGF Type 2 / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics; D014779:Virus Replication
D056827:Endosomal Sorting Complexes Required for Transport; D006388:Hemagglutinins; D007976:Leupeptins; D008164:Luminescent Proteins; D017527:Receptor, IGF Type 2; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin; C107528:VPS4 protein, S cerevisiae; C067713:lactacystin; D049452:Green Fluorescent Proteins; D000251:Adenosine Triphosphatases; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde; D000111:Acetylcysteine