title: A Polymerase mechanism-based strategy for viral attenuation and vaccine development.
abstract: Live, attenuated vaccines have prevented morbidity and mortality associated with myriad viral pathogens. Development of live, attenuated vaccines has traditionally relied on empirical methods, such as growth in nonhuman cells. These approaches require substantial time and expense to identify vaccine candidates and to determine their mechanisms of attenuation. With these constraints, at least a decade is required for approval of a live, attenuated vaccine for use in humans. We recently reported the discovery of an active site lysine residue that contributes to the catalytic efficiency of all nucleic acid polymerases (Castro, C., Smidansky, E. D., Arnold, J. J., Maksimchuk, K. R., Moustafa, I., Uchida, A., Götte, M., Konigsberg, W., and Cameron, C. E. (2009) Nat. Struct. Mol. Biol. 16, 212-218). Here we use a model RNA virus and its polymerase to show that mutation of this residue from lysine to arginine produces an attenuated virus that is genetically stable and elicits a protective immune response. Given the conservation of this residue in all viral polymerases, this study suggests that a universal, mechanism-based strategy may exist for viral attenuation and vaccine development.
authors.affiliation: Department of Biochemistry and Molecular Biology, The Pennsylvania State University,University Park, Pennsylvania 16802, USA.
authors.forename: Spencer A
authors.initials: SA
authors.lastname: Weeks
authors.forename: Cheri A
authors.initials: CA
authors.lastname: Lee
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhao
authors.forename: Eric D
authors.initials: ED
authors.lastname: Smidansky
authors.forename: Avery
authors.initials: A
authors.lastname: August
authors.forename: Jamie J
authors.initials: JJ
authors.lastname: Arnold
authors.forename: Craig E
authors.initials: CE
authors.lastname: Cameron
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000737:chemistry; D001709:Biotechnology / Q000379:methods; D002384:Catalysis; D020134:Catalytic Domain; D016895:Culture Media, Serum-Free / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008239:Lysine / Q000737:chemistry; D051379:Mice; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014613:Vaccines, Attenuated / Q000235:genetics / Q000276:immunology*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D016895:Culture Media, Serum-Free; D004279:DNA, Viral; D014613:Vaccines, Attenuated; D014764:Viral Proteins; D001120:Arginine; D004259:DNA-Directed DNA Polymerase; D008239:Lysine