Utilization of sialic acid as a coreceptor enhances reovirus attachment by multistep adhesion strengthening.
Many serotype 3 reoviruses bind to two different host cell molecules, sialic acid and an unidentified protein, using discrete receptor-binding domains in viral attachment protein, final sigma1. To determine mechanisms by which these receptor-binding events cooperate to mediate cell attachment, we generated isogenic reovirus strains that differ in the capacity to bind sialic acid. Strain SA+, but not SA-, bound specifically to sialic acid on a biosensor chip with nanomolar avidity. SA+ displayed 5-fold higher avidity for HeLa cells when compared with SA-, although both strains recognized the same proteinaceous receptor. Increased avidity of SA+ binding was mediated by increased k(on). Neuraminidase treatment to remove cell-surface sialic acid decreased the k(on) of SA+ to that of SA-. Increased k(on) of SA+ enhanced an infectious attachment process, since SA+ was 50-100-fold more efficient than SA- at infecting HeLa cells in a kinetic fluorescent focus assay. Sialic acid binding was operant early during SA+ attachment, since the capacity of soluble sialyllactose to inhibit infection decreased rapidly during the first 20 min of adsorption. These results indicate that reovirus binding to sialic acid enhances virus infection through adhesion of virus to the cell surface where access to a proteinaceous receptor is thermodynamically favored.
Barton
E S
ES
Department of Microbiology and Immunology, Elizabeth B. Lamb Center for Pediatric Research, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2581, USA.
Connolly
J L
JL
Forrest
J C
JC
Chappell
J D
JD
Dermody
T S
TS
D000818:Animals; D015374:Biosensing Techniques; D002460:Cell Line; D006801:Humans; D007700:Kinetics; D008561:Membrane Fusion; D051379:Mice; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D011485:Protein Binding; D011991:Receptors, Virus / Q000378:metabolism*; D012087:Reoviridae / Q000502:physiology*; D020349:Surface Plasmon Resonance
D011991:Receptors, Virus; D019158:N-Acetylneuraminic Acid