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Crystal structure of the C-terminal domain of splicing factor Prp8 carrying retinitis pigmentosa mutants

Authors

  • Lingdi Zhang,

    1. Department of Biochemistry and Molecular Genetics, University of Colorado at Denver and Health Sciences Center, Aurora, Colorado 80045, USA
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    • These authors contributed equally to this work.

  • Jingping Shen,

    1. Department of Biochemistry and Molecular Genetics, University of Colorado at Denver and Health Sciences Center, Aurora, Colorado 80045, USA
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    • These authors contributed equally to this work.

  • Michael T. Guarnieri,

    1. Department of Biochemistry and Molecular Genetics, University of Colorado at Denver and Health Sciences Center, Aurora, Colorado 80045, USA
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  • Annie Heroux,

    1. Biology Department, Brookhaven National Laboratory, Upton, New York 11973-5000, USA
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  • Kui Yang,

    1. Department of Biochemistry and Molecular Genetics, University of Colorado at Denver and Health Sciences Center, Aurora, Colorado 80045, USA
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  • Rui Zhao

    Corresponding author
    1. Department of Biochemistry and Molecular Genetics, University of Colorado at Denver and Health Sciences Center, Aurora, Colorado 80045, USA
    • M.S. 8101, P.O. Box 6511, Aurora, CO 80045, USA; fax: (303) 724-3215.
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Abstract

Prp8 is a critical pre-mRNA splicing factor. Prp8 is proposed to help form and stabilize the spliceosome catalytic core and to be an important regulator of spliceosome activation. Mutations in human Prp8 (hPrp8) cause a severe form of the genetic disorder retinitis pigmentosa, RP13. Understanding the molecular mechanism of Prp8's function in pre-mRNA splicing and RP13 has been hindered by its large size (over 2000 amino acids) and remarkably low-sequence similarity with other proteins. Here we present the crystal structure of the C-terminal domain (the last 273 residues) of Caenorhabditis elegans Prp8 (cPrp8). The core of the C-terminal domain is an α/β structure that forms the MPN (Mpr1, Pad1 N-terminal) fold but without Zn2+ coordination. We propose that the C-terminal domain is a protein interaction domain instead of a Zn2+-dependent metalloenzyme as proposed for some MPN proteins. Mapping of RP13 mutants on the Prp8 structure suggests that these residues constitute a binding surface between Prp8 and other partner(s), and the disruption of this interaction provides a plausible molecular mechanism for RP13.

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