title: Probing FinO-FinP RNA interactions by site-directed protein-RNA crosslinking and gelFRET.
abstract: The conjugative transfer of F-plasmids is repressed by a two-component system, which consists of the antisense RNA FinP and the protein FinO. FinO binds FinP, protecting it from endonucleolytic degradation and facilitating duplex formation between FinP and its complementary RNA. Here we present the results of site-specific protein-RNA cross-linking and gel-based fluorescence resonance energy transfer (gelFRET) experiments used to probe the structure of a complex of FinO bound to an RNA target consisting of a duplex with 5' and 3' single-stranded tails. The crosslinking experiments reveal that an extensive, largely positively charged surface on FinO contacts RNA. The gelFRET measurements indicate that the 5' single-stranded tail of the RNA is in closer contact with much of the protein than the distal, blunt end of the RNA duplex. These data suggest that significant conformational adjustments in the protein and/or the RNA accompany complex formation.
authors.affiliation: Department of Biochemistry, University of Alberta, Edmonton, Canada.
authors.forename: Alexandru F
authors.initials: AF
authors.lastname: Ghetu
authors.forename: David C
authors.initials: DC
authors.lastname: Arthur
authors.forename: Tom K
authors.initials: TK
authors.lastname: Kerppola
authors.forename: J N Mark
authors.initials: JN
authors.lastname: Glover
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D003432:Cross-Linking Reagents / Q000737:chemistry*; D017931:DNA Primers; D029968:Escherichia coli Proteins; D005144:F Factor; D015335:Molecular Probes; D017354:Point Mutation; D012313:RNA / Q000737:chemistry*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D012097:Repressor Proteins
chemical_list: D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D017931:DNA Primers; D029968:Escherichia coli Proteins; D015335:Molecular Probes; D016601:RNA-Binding Proteins; D012097:Repressor Proteins; C074275:FinO protein, E coli; D012313:RNA