title: Association of ebola virus matrix protein VP40 with microtubules.
abstract: Viruses exploit a variety of cellular components to complete their life cycles, and it has become increasingly clear that use of host cell microtubules is a vital part of the infection process for many viruses. A variety of viral proteins have been identified that interact with microtubules, either directly or via a microtubule-associated motor protein. Here, we report that Ebola virus associates with microtubules via the matrix protein VP40. When transfected into mammalian cells, a fraction of VP40 colocalized with microtubule bundles and VP40 coimmunoprecipitated with tubulin. The degree of colocalization and microtubule bundling in cells was markedly intensified by truncation of the C terminus to a length of 317 amino acids. Further truncation to 308 or fewer amino acids abolished the association with microtubules. Both the full-length and the 317-amino-acid truncation mutant stabilized microtubules against depolymerization with nocodazole. Direct physical interaction between purified VP40 and tubulin proteins was demonstrated in vitro. A region of moderate homology to the tubulin binding motif of the microtubule-associated protein MAP2 was identified in VP40. Deleting this region resulted in loss of microtubule stabilization against drug-induced depolymerization. The presence of VP40-associated microtubules in cells continuously treated with nocodazole suggested that VP40 promotes tubulin polymerization. Using an in vitro polymerization assay, we demonstrated that VP40 directly enhances tubulin polymerization without any cellular mediators. These results suggest that microtubules may play an important role in the Ebola virus life cycle and potentially provide a novel target for therapeutic intervention against this highly pathogenic virus.
authors.affiliation: USAMRIID, 1425 Porter St., Frederick, MD 21702, USA.
authors.forename: Gordon
authors.initials: G
authors.lastname: Ruthel
authors.forename: Gretchen L
authors.initials: GL
authors.lastname: Demmin
authors.forename: George
authors.initials: G
authors.lastname: Kallstrom
authors.forename: Melodi P
authors.initials: MP
authors.lastname: Javid
authors.forename: Shirin S
authors.initials: SS
authors.lastname: Badie
authors.forename: Amy B
authors.initials: AB
authors.lastname: Will
authors.forename: Timothy
authors.initials: T
authors.lastname: Nelle
authors.forename: Rowena
authors.initials: R
authors.lastname: Schokman
authors.forename: Tam L
authors.initials: TL
authors.lastname: Nguyen
authors.forename: John H
authors.initials: JH
authors.lastname: Carra
authors.forename: Sina
authors.initials: S
authors.lastname: Bavari
authors.forename: M Javad
authors.initials: MJ
authors.lastname: Aman
mesh_terms: D002460:Cell Line; D029043:Ebolavirus / Q000235:genetics*; D006801:Humans; D007700:Kinetics; D008870:Microtubules / Q000821:virology*; D014162:Transfection; D014404:Tubulin / Q000378:metabolism; D014763:Viral Matrix Proteins / Q000378:metabolism*
chemical_list: D014404:Tubulin; C080981:VP40 protein, virus; D014763:Viral Matrix Proteins