title: Assembly of core helices and rapid tertiary folding of a small bacterial group I ribozyme.
abstract: Compact but non-native intermediates have been implicated in the hierarchical folding of several large RNAs, but there is little information on their structure. In this article, ribonuclease and hydroxyl radical cleavage protection assays showed that base pairing of core helices stabilize a compact state of a small group I ribozyme from Azoarcus pre-tRNA(ile). Base pairing of the ribozyme core requires 10-fold less Mg(2+) than stable tertiary interactions, indicating that assembly of helices in the catalytic core represents a distinct phase that precedes the formation of native tertiary structure. Tertiary folding occurs in <100 ms at 37 degrees C. Such rapid folding is unprecedented among group I ribozymes and illustrates the association between structural complexity and folding time. A 3D model of the Azoarcus ribozyme was constructed by identifying homologous sequence motifs in rRNA. The model reveals distinct structural features, such as a large interface between the P4-P6 and P3-P9 domains, that may explain the unusual stability of the Azoarcus ribozyme and the cooperativity of folding.
authors.affiliation: T. C. Jenkins Department of Biophysics, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218, USA.
authors.forename: Prashanth
authors.initials: P
authors.lastname: Rangan
authors.forename: Benoît
authors.initials: B
authors.lastname: Masquida
authors.forename: Eric
authors.initials: E
authors.lastname: Westhof
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Woodson
mesh_terms: D020627:Azoarcus / Q000201:enzymology*; D001483:Base Sequence; D007438:Introns; D008274:Magnesium / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D016337:RNA, Catalytic / Q000737:chemistry* / Q000235:genetics
chemical_list: D016337:RNA, Catalytic; D008274:Magnesium