title: Proteins associated with SF3a60 in T. brucei.
abstract: Trypanosoma brucei relies on Spliced leader trans splicing to generate functional messenger RNAs. Trans splicing joins the specialized SL exon from the SL RNA to pre-mRNAs and is mediated by the trans-spliceosome, which is made up of small nuclear ribonucleoprotein particles and non-snRNP factors. Although the trans spliceosome is essential for trypanosomatid gene expression, not all spliceosomal protein factors are known and of these, only a few are completely characterized. In this study, we have characterized the trypanosome Splicing Factor, SF3a60, the only currently annotated SF3a component. As expected, epitope-tagged SF3a60 localizes in the trypanosome nucleus. SF3a60 is essential for cell viability but its depletion seem to have no detectable effect on trans-splicing. In addition, we used SF3a60 as bait in a Yeast-2-hybrid system screen and identified its interacting protein factors. The interactions with SF3a120, SF3a66 and SAP130 were confirmed by tandem affinity purification and mass spectrometry.
authors.affiliation: Laboratorio de Biología Molecular de la Enfermedad de Chagas, Instituto de Investigacíones en Ingeniería Genética y Biología Molecular (INGEBI), Buenos Aires, Argentina; Molecular Biology and Biotechnology Department, International Center for Insect Physiology and Ecology (ICIPE), Nairobi, Kenya; Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Heidelberg, Germany.
authors.forename: Benson
authors.initials: B
authors.lastname: Nyambega
authors.affiliation: Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Heidelberg, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Helbig
authors.affiliation: Molecular Biology and Biotechnology Department, International Center for Insect Physiology and Ecology (ICIPE), Nairobi, Kenya.
authors.forename: Daniel K
authors.initials: DK
authors.lastname: Masiga
authors.affiliation: Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), Heidelberg, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Clayton
authors.affiliation: Laboratorio de Biología Molecular de la Enfermedad de Chagas, Instituto de Investigacíones en Ingeniería Genética y Biología Molecular (INGEBI), Buenos Aires, Argentina.
authors.forename: Mariano J
authors.initials: MJ
authors.lastname: Levin
mesh_terms: D000595:Amino Acid Sequence; D002467:Cell Nucleus / Q000378:metabolism; D002470:Cell Survival; D002846:Chromatography, Affinity; D017124:Conserved Sequence; D020868:Gene Silencing; D008969:Molecular Sequence Data; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D015800:Protozoan Proteins / Q000737:chemistry / Q000378:metabolism*; D017413:Ribonucleoprotein, U2 Small Nuclear / Q000737:chemistry / Q000378:metabolism*; D020040:Trans-Splicing; D014346:Trypanosoma brucei brucei / Q000166:cytology / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
chemical_list: D015800:Protozoan Proteins; D017413:Ribonucleoprotein, U2 Small Nuclear; C088049:splicing factor 3a