title: Ebola virus enters host cells by macropinocytosis and clathrin-mediated endocytosis.
abstract: Virus entry into host cells is the first step of infection and a crucial determinant of pathogenicity. Here we show that Ebola virus-like particles (EBOV-VLPs) composed of the glycoprotein GP(1,2) and the matrix protein VP40 use macropinocytosis and clathrin-mediated endocytosis to enter cells. EBOV-VLPs applied to host cells induced actin-driven ruffling and enhanced FITC-dextran uptake, which indicated macropinocytosis as the main entry mechanism. This was further supported by inhibition of entry through inhibitors of actin polymerization (latrunculin A), Na(+)/H(+)-exchanger (EIPA), and PI3-kinase (wortmannin). A fraction of EBOV-VLPs, however, colocalized with clathrin heavy chain (CHC), and VLP uptake was reduced by CHC small interfering RNA transfection and expression of the dominant negative dynamin II-K44A mutant. In contrast, we found no evidence that EBOV-VLPs enter cells via caveolae. This work identifies macropinocytosis as the major, and clathrin-dependent endocytosis as an alternative, entry route for EBOV particles. Therefore, EBOV seems to utilize different entry pathways depending on both cell type and virus particle size.
authors: Institute of Anatomy and Vascular Biology, Westfälische Wilhelms University Muenster, Vesaliusweg, Germany.
authors: Paulina
authors: P
authors: Aleksandrowicz
authors: Andrea
authors: A
authors: Marzi
authors: Nadine
authors: N
authors: Biedenkopf
authors: Nadine
authors: N
authors: Beimforde
authors: Stephan
authors: S
authors: Becker
authors: Thomas
authors: T
authors: Hoenen
authors: Heinz
authors: H
authors: Feldmann
authors: Hans-Joachim
authors: HJ
authors: Schnittler
mesh_terms: D000199:Actins / Q000502:physiology; D000584:Amiloride / Q000031:analogs & derivatives / Q000494:pharmacology; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D051242:Caveolin 1 / Q000378:metabolism; D002460:Cell Line; D002966:Clathrin / Q000502:physiology*; D029043:Ebolavirus / Q000502:physiology*; D004705:Endocytosis / Q000502:physiology*; D004847:Epithelial Cells / Q000378:metabolism / Q000821:virology*; D020868:Gene Silencing; D054884:Host-Pathogen Interactions; D006801:Humans; D034741:RNA, Small Interfering; D015398:Signal Transduction; D053778:Thiazolidines / Q000494:pharmacology; D053586:Virus Internalization; D014779:Virus Replication; D000077191:Wortmannin
chemical_list: D000199:Actins; D000730:Androstadienes; D019086:Bridged Bicyclo Compounds, Heterocyclic; D051242:Caveolin 1; D002966:Clathrin; D034741:RNA, Small Interfering; D053778:Thiazolidines; D000584:Amiloride; C037067:latrunculin A; C039614:ethylisopropylamiloride; D000077191:Wortmannin