title: Flexibility of the adenovirus fiber is required for efficient receptor interaction.
abstract: The adenovirus (Ad) fiber protein mediates Ad binding to the coxsackievirus and Ad receptor (CAR) and is thus a major determinant of viral tropism. The fiber contains three domains: an N-terminal tail that anchors the fiber to the viral capsid, a central shaft region of variable length and flexibility, and a C-terminal knob domain that binds to cell receptors. Ad type 37 (Ad37), a subgroup D virus associated with severe ocular infections, is unable to use CAR efficiently to infect host cells, despite containing a CAR binding site in its fiber knob. We hypothesized that the relatively short, inflexible Ad37 fiber protein restricts interactions with CAR at the cell surface. To test this hypothesis, we analyzed the infectivity and binding of recombinant Ad particles containing modified Ad37 or Ad5 fiber proteins. Ad5 particles equipped with a truncated Ad5 fiber or with a chimeric fiber protein comprised of the Ad5 knob fused to the short, rigid Ad37 shaft domain had significantly reduced infectivity and attachment. In contrast, placing the Ad37 knob onto the long, flexible Ad5 shaft allowed CAR-dependent virus infection and cell attachment, demonstrating the importance of the shaft domain in receptor usage. Increasing fiber rigidity by substituting the predicted flexibility modules in the Ad5 shaft with the corresponding regions of the rigid Ad37 fiber dramatically reduced both virus infection and cell attachment. Cryoelectron microscopy (cryo-EM) single-particle analysis demonstrated the increased rigidity of this chimeric fiber. These studies demonstrate that both length and flexibility of the fiber shaft regulate CAR interaction and provide a molecular explanation for the use of alternative receptors by subgroup D Ad with ocular tropism. We present a molecular model for Ad-CAR interactions at the cell surface that explains the significance of fiber flexibility in cell attachment.
authors.affiliation: Department of Immunology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Eugene
authors.initials: E
authors.lastname: Wu
authors.forename: Lars
authors.initials: L
authors.lastname: Pache
authors.forename: Dan J
authors.initials: DJ
authors.lastname: Von Seggern
authors.forename: Tina-Marie
authors.initials: TM
authors.lastname: Mullen
authors.forename: Yeshi
authors.initials: Y
authors.lastname: Mikyas
authors.forename: Phoebe L
authors.initials: PL
authors.lastname: Stewart
authors.forename: Glen R
authors.initials: GR
authors.lastname: Nemerow
mesh_terms: D000256:Adenoviridae / Q000502:physiology* / Q000648:ultrastructure; D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D020285:Cryoelectron Microscopy; D017931:DNA Primers; D006801:Humans; D008969:Molecular Sequence Data; D011991:Receptors, Virus / Q000737:chemistry / Q000502:physiology*; D017386:Sequence Homology, Amino Acid
chemical_list: D017931:DNA Primers; D011991:Receptors, Virus; C085301:adenovirus receptor