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Spliceosomal Proteomics in Trypanosoma brucei Reveal New RNA Splicing Factors

dc.contributor.authorAmbrosio, Daniela Luz [UNESP]
dc.contributor.authorLee, Ju Huck
dc.contributor.authorPanigrahi, Aswini K.
dc.contributor.authorNguyen, Tu Ngoc
dc.contributor.authorCicarelli, Regina Maria Barretto [UNESP]
dc.contributor.authorGuenzl, Arthur
dc.contributor.institutionUniv Connecticut
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionSeattle Biomed Res Inst
dc.date.accessioned2014-05-20T15:34:24Z
dc.date.available2014-05-20T15:34:24Z
dc.date.issued2009-07-01
dc.description.abstractIn trypanosomatid parasites, spliced leader (SL) trans splicing is an essential nuclear mRNA maturation step which caps mRNAs posttranscriptionally and, in conjunction with polyadenylation, resolves individual mRNAs from polycistronic precursors. While all trypanosomatid mRNAs are trans spliced, intron removal by cis splicing is extremely rare and predicted to occur in only four pre-mRNAs. trans-and cis-splicing reactions are carried out by the spliceosome, which consists of U-rich small nuclear ribonucleoprotein particles (U snRNPs) and of non-snRNP factors. Mammalian and yeast spliceosome complexes are well characterized and found to be associated with up to 170 proteins. Despite the central importance of trans splicing in trypanosomatid gene expression, only the core RNP proteins and a few snRNP-specific proteins are known. To characterize the trypanosome spliceosomal protein repertoire, we conducted a proteomic analysis by tagging and tandem affinity-purifying the canonical core RNP protein SmD1 in Trypanosoma brucei and by identifying copurified proteins by mass spectrometry. The set of 47 identified proteins harbored nearly all spliceosomal snRNP factors characterized in trypanosomes thus far and 21 proteins lacking a specific annotation. A bioinformatic analysis combined with protein pull-down assays and immunofluorescence microscopy identified 10 divergent orthologues of known splicing factors, including the missing U1-specific protein U1A. In addition, a novel U5-specific, and, as we show, an essential splicing factor was identified that shares a short, highly conserved N-terminal domain with the yeast protein Cwc21p and was thus tentatively named U5-Cwc21. Together, these data strongly indicate that most of the identified proteins are components of the spliceosome.en
dc.description.affiliationUniv Connecticut, Ctr Hlth, Dept Genet & Dev Biol, Farmington, CT 06030 USA
dc.description.affiliationUniv Connecticut, Ctr Hlth, Dept Mol Microbial & Struct Biol, Farmington, CT 06030 USA
dc.description.affiliationUniv Estadual Paulista, Fac Ciencias Farmaceut, Dept Ciencias Biol, São Paulo, Brazil
dc.description.affiliationSeattle Biomed Res Inst, Seattle, WA 98109 USA
dc.description.affiliationUnespUniv Estadual Paulista, Fac Ciencias Farmaceut, Dept Ciencias Biol, São Paulo, Brazil
dc.description.sponsorshipNational Institutes of Health
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.description.sponsorshipKorea Science and Engineering Foundation (KOSEF)
dc.description.sponsorshipIdNIH: AI059377
dc.description.sponsorshipIdFAPESP: 07/07476-0
dc.description.sponsorshipIdCAPES: BEX 1338/06-4
dc.description.sponsorshipIdKorea Science and Engineering Foundation: C00093
dc.format.extent990-1000
dc.identifierhttp://dx.doi.org/10.1128/EC.00075-09
dc.identifier.citationEukaryotic Cell. Washington: Amer Soc Microbiology, v. 8, n. 7, p. 990-1000, 2009.
dc.identifier.doi10.1128/EC.00075-09
dc.identifier.fileWOS000268760600007.pdf
dc.identifier.issn1535-9778
dc.identifier.urihttp://hdl.handle.net/11449/42537
dc.identifier.wosWOS:000268760600007
dc.language.isoeng
dc.publisherAmer Soc Microbiology
dc.relation.ispartofEukaryotic Cell
dc.relation.ispartofsjr1,603
dc.rights.accessRightsAcesso abertopt
dc.sourceWeb of Science
dc.titleSpliceosomal Proteomics in Trypanosoma brucei Reveal New RNA Splicing Factorsen
dc.typeArtigopt
dcterms.licensehttp://journals.asm.org/site/misc/ASM_Author_Statement.xhtml
dcterms.rightsHolderAmer Soc Microbiology
dspace.entity.typePublication
relation.isDepartmentOfPublication5004bcab-94af-4939-b980-091ae9d0a19e
relation.isDepartmentOfPublication.latestForDiscovery5004bcab-94af-4939-b980-091ae9d0a19e
relation.isOrgUnitOfPublication95697b0b-8977-4af6-88d5-c29c80b5ee92
relation.isOrgUnitOfPublication.latestForDiscovery95697b0b-8977-4af6-88d5-c29c80b5ee92
unesp.campusUniversidade Estadual Paulista (UNESP), Faculdade de Ciências Farmacêuticas, Araraquarapt
unesp.departmentCiências Biológicas - FCFpt

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