title: The spliceosomal PRP19 complex of trypanosomes.
abstract: In trypanosomes, mRNAs are processed by spliced leader (SL) trans splicing, in which a capped SL, derived from SL RNA, is spliced onto the 5' end of each mRNA. This process is mediated by the spliceosome, a large and dynamic RNA-protein machinery consisting of small nuclear ribonucleoproteins (snRNPs) and non-snRNP proteins. Due to early evolutionary divergence, the amino acid sequences of trypanosome splicing factors exhibit limited similarity to those of their eukaryotic orthologs making their bioinformatic identification challenging. Most of the ~ 60 protein components that have been characterized thus far are snRNP proteins because, in contrast to individual snRNPs, purification of intact spliceosomes has not been achieved yet. Here, we characterize the non-snRNP PRP19 complex of Trypanosoma brucei. We identified a complex that contained the core subunits PRP19, CDC5, PRL1, and SPF27, as well as PRP17, SKIP and PPIL1. Three of these proteins were newly annotated. The PRP19 complex was associated primarily with the activated spliceosome and, accordingly, SPF27 silencing blocked the first splicing step. Interestingly, SPF27 silencing caused an accumulation of SL RNA with a hypomethylated cap that closely resembled the defect observed previously upon depletion of the cyclin-dependent kinase CRK9, indicating that both proteins may function in spliceosome activation.
authors.affiliation: Department of Genetics and Genome Sciences, University of Connecticut Health Center, 400 Farmington Avenue, Farmington, CT, 06030-6403, USA.
authors.forename: Daniela L
authors.initials: DL
authors.lastname: Ambrósio
authors.forename: Nitika
authors.initials: N
authors.lastname: Badjatia
authors.forename: Arthur
authors.initials: A
authors.lastname: Günzl
mesh_terms: D000595:Amino Acid Sequence; D005455:Fluorescent Antibody Technique; D013058:Mass Spectrometry; D046912:Multiprotein Complexes / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D012326:RNA Splicing; D016053:RNA, Protozoan / Q000378:metabolism; D012342:RNA, Small Nuclear / Q000378:metabolism; D017411:Ribonucleoproteins, Small Nuclear / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D017381:Spliceosomes; D014346:Trypanosoma brucei brucei / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism
chemical_list: D046912:Multiprotein Complexes; D021122:Protein Subunits; D015800:Protozoan Proteins; D016053:RNA, Protozoan; D012342:RNA, Small Nuclear; D017411:Ribonucleoproteins, Small Nuclear