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| Subject Categories: RNA | Genomic & Computational Biology |
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| The EMBO Journal
(2004) 23, 4847–4856, doi:10.1038/sj.emboj.7600482 Published online 25 November 2004 |
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| Proteomic analysis identifies a new complex required for nuclear pre-mRNA retention and splicing |
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| Andrzej Dziembowski1, Ana-Paula Ventura2, Berthold Rutz3, 4, Friederike Caspary3, 5, Céline Faux1, Frédéric Halgand2, Olivier Laprévote2 and Bertrand Séraphin1, 3 |
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1 Centre de Génétique Moléculaire, Avenue de la Terrasse, Gif sur Yvette, France 2 ICSN, Avenue de la Terrasse, Gif sur Yvette, France 3 EMBL, Heidelberg, Germany To whom correspondence should be addressed Bertrand Séraphin, Centre de Génétique Moléculaire, CNRS UPR2167, Avenue de la Terrasse, 91198 Gif sur Yvette, France. Tel.: +33 1 69 82 38 84; Fax: +33 1 69 82 38 77; E-mail: seraphin@cgm.cnrs-gif.fr 4 Present address: EPO, Landsbergerstr. 30, 80339 München, Germany 5 Present address: MediGene, Lochhamerstr. 11, 82152 Martinsried, Germany Received 29 June 2004; Accepted 21 October 2004; Published online 25 November 2004. |
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| Abstract |
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| Using the proteomic tandem affinity purification (TAP) method, we have purified the Saccharomyces cerevisie U2 snRNP-associated splicing factors SF3a and SF3b. While SF3a purification revealed only the expected subunits Prp9p, Prp11p and Prp21p, yeast SF3b was found to contain only six subunits, including previously known components (Rse1p, Hsh155p, Cus1p, Hsh49p), the recently identified Rds3p factor and a new small essential protein (Ysf3p) encoded by an unpredicted split ORF in the yeast genome. Surprisingly, Snu17p, the proposed yeast orthologue of the seventh human SF3b subunit, p14, was not found in the yeast complex. TAP purification revealed that Snu17p, together with Bud13p and a newly identified factor, Pml1p/Ylr016c, form a novel trimeric complex. Subunits of this complex were not essential for viability. However, they are required for efficient splicing in vitro and in vivo. Furthermore, inactivation of this complex causes pre-mRNA leakage from the nucleus. The corresponding complex was named pre-mRNA REtention and Splicing (RES). The presence of RES subunit homologues in numerous eukaryotes suggests that its function is evolutionarily conserved. |
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| Keywords: RES, SF3a, SF3b, tandem affinity purification (TAP), U2 snRNP |
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