title: NMR structure of stem-loop D from human rhinovirus-14.
abstract: The 5'-cloverleaf of the picornavirus RNA genome is essential for the assembly of a ribonucleoprotein replication complex. Stem-loop D (SLD) of the cloverleaf is the recognition site for the multifunctional viral protein 3Cpro. This protein is the principal viral protease, and its interaction with SLD also helps to position the viral RNA-dependent RNA polymerase (3Dpol) for replication. Human rhinovirus-14 (HRV-14) is distinct from the majority of picornaviruses in that its SLD forms a cUAUg triloop instead of the more common uYACGg tetraloop. This difference appears to be functionally significant, as 3Cpro from tetraloop-containing viruses cannot bind the HRV-14 SLD. We have determined the solution structure of the HRV-14 SLD using NMR spectroscopy. The structure is predominantly an A-form helix, but with a central pyrimidine-pyrimidine base-paired region and a significantly widened major groove. The stabilizing hydrogen bonding present in the uYACGg tetraloop was not found in the cUAUg triloop. However, the triloop uses different structural elements to present a largely similar surface: sequence and underlying architecture are not conserved, but key aspects of the surface structure are. Important structural differences do exist, though, and may account for the observed cross-isotype binding specificities between 3Cpro and SLD.
authors.affiliation: Institute of Fundamental Sciences, Massey University, Palmerston North, New Zealand.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Headey
authors.forename: He
authors.initials: H
authors.lastname: Huang
authors.forename: Jolyon K
authors.initials: JK
authors.lastname: Claridge
authors.forename: Giselle A
authors.initials: GA
authors.lastname: Soares
authors.forename: Kaushik
authors.initials: K
authors.lastname: Dutta
authors.forename: Martin
authors.initials: M
authors.lastname: Schwalbe
authors.forename: Daiwen
authors.initials: D
authors.lastname: Yang
authors.forename: Steven M
authors.initials: SM
authors.lastname: Pascal
mesh_terms: D002241:Carbohydrates / Q000737:chemistry; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D009690:Nucleic Acid Conformation; D011743:Pyrimidines / Q000737:chemistry; D012367:RNA, Viral / Q000737:chemistry*; D012229:Rhinovirus / Q000235:genetics*
chemical_list: D002241:Carbohydrates; D011743:Pyrimidines; D012367:RNA, Viral; C030986:pyrimidine