title: The Prp19 WD40 domain contains a conserved protein interaction region essential for its function.
abstract: Prp19 is a member of the WD40 repeat family of E3 ubiquitin ligases and a conserved eukaryotic RNA splicing factor essential for activation and stabilization of the spliceosome. To understand the role of the WD40 repeat domain of Prp19 we have determined its structure using X-ray crystallography. The domain has a distorted seven bladed WD40 architecture with significant asymmetry due to irregular packing of blades one and seven into the core of the WD40 domain. Structure-based mutagenesis identified a highly conserved surface centered around blade five that is required for the physical interaction between Prp19 and Cwc2, another essential splicing factor. This region is found to be required for Prp19 function and yeast viability. Experiments in vitro and in vivo demonstrate that two molecules of Cwc2 bind to the Prp19 tetramer. These coupled structural and functional studies provide a model for the functional architecture of Prp19.
authors.affiliation: Department of Biochemistry, Vanderbilt University, Nashville, TN 37232, USA. craig.vanderkooi@uky.edu
authors.forename: Craig W
authors.initials: CW
authors.lastname: Vander Kooi
authors.forename: Liping
authors.initials: L
authors.lastname: Ren
authors.forename: Ping
authors.initials: P
authors.lastname: Xu
authors.forename: Melanie D
authors.initials: MD
authors.lastname: Ohi
authors.forename: Kathleen L
authors.initials: KL
authors.lastname: Gould
authors.forename: Walter J
authors.initials: WJ
authors.lastname: Chazin
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D018360:Crystallography, X-Ray; D056890:Eukaryota; D016296:Mutagenesis; D017434:Protein Structure, Tertiary / Q000235:genetics; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D012326:RNA Splicing; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D017381:Spliceosomes / Q000235:genetics / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014965:X-Rays
chemical_list: D011506:Proteins; D044767:Ubiquitin-Protein Ligases
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