title: Evolutionarily divergent spliceosomal snRNAs and a conserved non-coding RNA processing motif in Giardia lamblia.
abstract: Non-coding RNAs (ncRNAs) have diverse essential biological functions in all organisms, and in eukaryotes, two such classes of ncRNAs are the small nucleolar (sno) and small nuclear (sn) RNAs. In this study, we have identified and characterized a collection of sno and snRNAs in Giardia lamblia, by exploiting our discovery of a conserved 12 nt RNA processing sequence motif found in the 3' end regions of a large number of G. lamblia ncRNA genes. RNA end mapping and other experiments indicate the motif serves to mediate ncRNA 3' end formation from mono- and di-cistronic RNA precursor transcripts. Remarkably, we find the motif is also utilized in the processing pathway of all four previously identified trans-spliced G. lamblia introns, revealing a common RNA processing pathway for ncRNAs and trans-spliced introns in this organism. Motif sequence conservation then allowed for the bioinformatic and experimental identification of additional G. lamblia ncRNAs, including new U1 and U6 spliceosomal snRNA candidates. The U6 snRNA candidate was then used as a tool to identity novel U2 and U4 snRNAs, based on predicted phylogenetically conserved snRNA-snRNA base-pairing interactions, from a set of previously identified G. lamblia ncRNAs without assigned function. The Giardia snRNAs retain the core features of spliceosomal snRNAs but are sufficiently evolutionarily divergent to explain the difficulties in their identification. Most intriguingly, all of these snRNAs show structural features diagnostic of U2-dependent/major and U12-dependent/minor spliceosomal snRNAs.
authors.affiliation: Alberta RNA Research and Training Institute, University of Lethbridge, Lethbridge, Alberta T1K 3M4, Canada.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Hudson
authors.forename: Ashley N
authors.initials: AN
authors.lastname: Moore
authors.forename: David
authors.initials: D
authors.lastname: Elniski
authors.forename: Joella
authors.initials: J
authors.lastname: Joseph
authors.forename: Janet
authors.initials: J
authors.lastname: Yee
authors.forename: Anthony G
authors.initials: AG
authors.lastname: Russell
mesh_terms: D001483:Base Sequence; D017124:Conserved Sequence; D019143:Evolution, Molecular; D016829:Giardia lamblia / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D059372:Nucleotide Motifs; D026721:RNA 3' End Processing; D012333:RNA, Messenger / Q000378:metabolism; D012342:RNA, Small Nuclear / Q000737:chemistry*; D020537:RNA, Small Nucleolar / Q000235:genetics; D022661:RNA, Untranslated / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D038621:Regulatory Sequences, Ribonucleic Acid; D017381:Spliceosomes / Q000737:chemistry*; D020040:Trans-Splicing
chemical_list: D012333:RNA, Messenger; D012342:RNA, Small Nuclear; D020537:RNA, Small Nucleolar; D022661:RNA, Untranslated; D038621:Regulatory Sequences, Ribonucleic Acid; C405483:U1 small nuclear RNA; C405484:U2 small nuclear RNA; C405482:U4 small nuclear RNA; C405486:U6 small nuclear RNA