The Prp19 WD40 domain contains a conserved protein interaction region essential for its function

Craig W. Vander Kooi; Liping Ren; Ping Xu; Melanie D. Ohi; Kathleen L. Gould; Walter J. Chazin

doi:10.1016/j.str.2010.02.015

The Prp19 WD40 domain contains a conserved protein interaction region essential for its function

Craig W. Vander Kooi, Liping Ren, Ping Xu, Melanie D. Ohi, Kathleen L. Gould, Walter J. Chazin

Research output: Contribution to journal › Article › peer-review

32 Scopus citations

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.

Original language	English
Pages (from-to)	584-593
Number of pages	10
Journal	Structure
Volume	18
Issue number	5
DOIs	https://doi.org/10.1016/j.str.2010.02.015
State	Published - May 2010

Bibliographical note

Funding Information:
We thank H. Bellamy for help in collection of X-ray diffraction data and M. Oldham and M. Newcomer for valuable advice and discussions. This work was supported by National Institutes of Health (NIH) training grant T32GM08320; by NIH operating grants P20RR020171 (C.W.V.K.), 1DP2OD004483 (M.D.O.), and R01GM075156 (W.J.C.); and by the Howard Hughes Medical Institute (K.L.G.).

Keywords

CELLBIO
PROTEINS
RNA

ASJC Scopus subject areas

Structural Biology
Molecular Biology

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10.1016/j.str.2010.02.015

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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.",

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author = "{Vander Kooi}, {Craig W.} and Liping Ren and Ping Xu and Ohi, {Melanie D.} and Gould, {Kathleen L.} and Chazin, {Walter J.}",

note = "Funding Information: We thank H. Bellamy for help in collection of X-ray diffraction data and M. Oldham and M. Newcomer for valuable advice and discussions. This work was supported by National Institutes of Health (NIH) training grant T32GM08320; by NIH operating grants P20RR020171 (C.W.V.K.), 1DP2OD004483 (M.D.O.), and R01GM075156 (W.J.C.); and by the Howard Hughes Medical Institute (K.L.G.). ",

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AU - Ohi, Melanie D.

AU - Gould, Kathleen L.

AU - Chazin, Walter J.

N1 - Funding Information: We thank H. Bellamy for help in collection of X-ray diffraction data and M. Oldham and M. Newcomer for valuable advice and discussions. This work was supported by National Institutes of Health (NIH) training grant T32GM08320; by NIH operating grants P20RR020171 (C.W.V.K.), 1DP2OD004483 (M.D.O.), and R01GM075156 (W.J.C.); and by the Howard Hughes Medical Institute (K.L.G.).

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N2 - 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.

AB - 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.

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The Prp19 WD40 domain contains a conserved protein interaction region essential for its function

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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