title: Rift Valley fever virus strain MP-12 enters mammalian host cells via caveola-mediated endocytosis.
abstract: Rift Valley fever virus (RVFV) is a zoonotic pathogen capable of causing serious morbidity and mortality in both humans and livestock. The lack of efficient countermeasure strategies, the potential for dispersion into new regions, and the pathogenesis in humans and livestock make RVFV a serious public health concern. The receptors, cellular factors, and entry pathways used by RVFV and other members of the family Bunyaviridae remain largely uncharacterized. Here we provide evidence that RVFV strain MP-12 uses dynamin-dependent caveola-mediated endocytosis for cell entry. Caveolae are lipid raft domains composed of caveolin (the main structural component), cholesterol, and sphingolipids. Caveola-mediated endocytosis is responsible for the uptake of a wide variety of host ligands, as well as bacteria, bacterial toxins, and a number of viruses. To determine the cellular entry mechanism of RVFV, we used small-molecule inhibitors, RNA interference (RNAi), and dominant negative (DN) protein expression to inhibit the major mammalian cell endocytic pathways. Inhibitors and RNAi specific for macropinocytosis and clathrin-mediated endocytosis had no effect on RVFV infection. In contrast, inhibitors of caveola-mediated endocytosis, and RNAi targeted to caveolin-1 and dynamin, drastically reduced RVFV infection in multiple cell lines. Expression of DN caveolin-1 also reduced RVFV infection significantly, while expression of DN EPS15, a protein required for the assembly of clathrin-coated pits, and DN PAK-1, an obligate mediator of macropinocytosis, had no significant impact on RVFV infection. These results together suggest that the primary mechanism of RVFV MP-12 uptake is dynamin-dependent, caveolin-1-mediated endocytosis.
authors.affiliation: Sandia National Laboratories, Livermore, California, USA.
authors.forename: Brooke
authors.initials: B
authors.lastname: Harmon
authors.forename: Benjamin R
authors.initials: BR
authors.lastname: Schudel
authors.forename: Dianna
authors.initials: D
authors.lastname: Maar
authors.forename: Carol
authors.initials: C
authors.lastname: Kozina
authors.forename: Tetsuro
authors.initials: T
authors.lastname: Ikegami
authors.forename: Chien-Te Kent
authors.initials: CT
authors.lastname: Tseng
authors.forename: Oscar A
authors.initials: OA
authors.lastname: Negrete
mesh_terms: D000818:Animals; D015153:Blotting, Western; D021941:Caveolae / Q000378:metabolism* / Q000502:physiology; D022461:Caveolins / Q000235:genetics; D002522:Chlorocebus aethiops; D004705:Endocytosis / Q000502:physiology*; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins; D057809:HEK293 Cells; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D012296:Rift Valley fever virus / Q000502:physiology*; D053586:Virus Internalization
chemical_list: D022461:Caveolins; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins