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Molecular and Cellular Biology, January 2005, p. 451-460, Vol. 25, No. 1
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.1.451-460.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Structural and Functional Analysis of Essential pre-mRNA Splicing Factor Prp19p

Melanie D. Ohi,1 Craig W. Vander Kooi,2,3 Joshua A. Rosenberg,4,5 Liping Ren,4,5 Justin P. Hirsch,1 Walter J. Chazin,2,3,6 Thomas Walz,1* and Kathleen L. Gould4,5*

Department of Cell Biology, Harvard Medical School, Boston, Massachusetts,1 Department of Biochemistry,2 Department of Physics,6 Center for Structural Biology,3 Howard Hughes Medical Institute,4 Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, Tennessee5

Received 18 July 2004/ Returned for modification 12 August 2004/ Accepted 28 September 2004

U-box-containing Prp19p is an integral component of the Prp19p-associated complex (the nineteen complex, or NTC) that is essential for activation of the spliceosome. Prp19p makes numerous protein-protein contacts with other NTC components and is required for NTC stability. Here we show that Prp19p forms a tetramer in vitro and in vivo and we map the domain required for its oligomerization to a central tetrameric coiled-coil. Biochemical and in vivo analyses are consistent with Prp19p tetramerization providing an interaction surface for a single copy of its binding partner, Cef1p. Electron microscopy showed that the isolated Prp19p tetramer is an elongated particle consisting of four globular WD40 domains held together by a central stalk consisting of four N-terminal U-boxes and four coiled-coils. These structural and functional data provide a basis for understanding the role of Prp19p as a key architectural component of the NTC.


* Corresponding author. Mailing address for Kathleen L. Gould: Department of Cell and Developmental Biology, Vanderbilt University, Nashville, TN 37232. Phone: (615) 343-9502. Fax: (615) 343-0723. E-mail: kathy.gould{at}vanderbilt.edu. Mailing address for Thomas Walz: Department of Cell Biology, Harvard Medical School, 240 Longwood Ave., Boston, MA 02115. Phone: (617) 432-4090. Fax: (617) 432-1144. E-mail: twalz{at}hms.harvard.edu.


Molecular and Cellular Biology, January 2005, p. 451-460, Vol. 25, No. 1
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.1.451-460.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.




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