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Molecular and Cellular Biology, December 2005, p. 10745-10754, Vol. 25, No. 24
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.24.10745-10754.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Interactions of the Yeast SF3b Splicing Factor{dagger}

Qiang Wang,1 Jin He,1 Bert Lynn,2 and Brian C. Rymond1*

Departments of Biology,1 Chemistry, University of Kentucky, Lexington, Kentucky 40506-02252

Received 15 June 2005/ Returned for modification 12 July 2005/ Accepted 21 September 2005

The U2 snRNP promotes prespliceosome assembly through interactions that minimally involve the branchpoint binding protein, Mud2p, and the pre-mRNA. We previously showed that seven proteins copurify with the yeast (Saccharomyces cerevisiae) SF3b U2 subcomplex that associates with the pre-mRNA branchpoint region: Rse1p, Hsh155p, Hsh49p, Cus1p, and Rds3p and unidentified subunits p10 and p17. Here proteomic and genetic studies identify Rcp10p as p10 and show that it contributes to SF3b stability and is necessary for normal cellular Cus1p accumulation and for U2 snRNP recruitment in splicing. Remarkably, only the final 53 amino acids of Rcp10p are essential. p17 is shown to be composed of two accessory splicing factors, Bud31p and Ist3p, the latter of which independently associates with the RES complex implicated in the nuclear pre-mRNA retention. A directed two-hybrid screen reveals a network of prospective interactions that includes previously unreported intra-SF3b contacts and SF3b interactions with the RES subunit Bud13p, the Prp5p DExD/H-box protein, Mud2p, and the late-acting nineteen complex. These data establish the concordance of yeast and mammalian SF3b complexes, implicate accessory splicing factors in U2 snRNP function, and support SF3b contribution from early pre-mRNP recognition to late steps in splicing.


* Corresponding author. Mailing address: Department of Biology, University of Kentucky, Lexington, KY 40506-0225. Phone: (859) 257-5530. Fax: (859) 257-1717. E-mail: rymond{at}uky.edu.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.


Molecular and Cellular Biology, December 2005, p. 10745-10754, Vol. 25, No. 24
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.24.10745-10754.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.




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