title: Splicing fidelity: DEAD/H-box ATPases as molecular clocks.
abstract: The spliceosome discriminates against suboptimal substrates, both during assembly and catalysis, thereby enhancing specificity during pre-mRNA splicing. Central to such fidelity mechanisms are a conserved subset of the DEAD- and DEAH-box ATPases, which belong to a superfamily of proteins that mediate RNP rearrangements in almost all RNA-dependent processes in the cell. Through an investigation of the mechanisms contributing to the specificity of 5' splice site cleavage, two related reports, one from our lab and the other from the Cheng lab, have provided insights into fidelity mechanisms utilized by the spliceosome. In our work, we found evidence for a kinetic proofreading mechanism in splicing in which the DEAH-box ATPase Prp16 discriminates against substrates undergoing slow 5' splice site cleavage. Additionally, our study revealed that discriminated substrates are discarded through a general spliceosome disassembly pathway, mediated by another DEAH-box ATPase Prp43. In their work, Tseng et al. described the underlying molecular events through which Prp16 discriminates against a splicing substrate during 5' splice site cleavage. Here, we present a synthesis of these two studies and, additionally, provide the first biochemical evidence for discrimination of a suboptimal splicing substrate just prior to 5' splice site cleavage. Together, these findings support a general mechanism for a ubiquitous superfamily of ATPases in enhancing specificity during RNA-dependent processes in the cell.
authors.affiliation: Department of Molecular Genetics and Cell Biology; The University of Chicago; Chicago, IL USA.
authors.forename: Prakash
authors.initials: P
authors.lastname: Koodathingal
authors.forename: Jonathan P
authors.initials: JP
authors.lastname: Staley
mesh_terms: D053487:DEAD-box RNA Helicases / Q000378:metabolism; D007438:Introns; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D022821:RNA Splice Sites; D012326:RNA Splicing / Q000502:physiology*; D017381:Spliceosomes / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D012322:RNA Precursors; D022821:RNA Splice Sites; D053487:DEAD-box RNA Helicases
keywords: ATPase; DEAD-box; DEAH-box; Prp16; Prp43; discard pathway; fidelity; kinetic proofreading; spliceosome; splicing