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Cloning of mDEAH9, a putative RNA helicase and mammalian homologue of Saccharomyces cerevisiae splicing factor Prp43

Gee, Sherry and Krauss, Sharon Wald and Miller, Estelle and Aoyagi, Kazuko and Arenas, Jaime and Conboy, John G. (1997) Cloning of mDEAH9, a putative RNA helicase and mammalian homologue of Saccharomyces cerevisiae splicing factor Prp43. Proceedings of the National Academy of Sciences of the United States of America, 94 (22). pp. 11803-11807. ISSN 0027-8424. PMCID PMC23596. doi:10.1073/pnas.94.22.11803.

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Yeast splicing factor Prp43, a DEAH box protein of the putative RNA helicase/RNA-dependent NTPase family, is a splicing factor that functions late in the pre-mRNA splicing pathway to facilitate spliceosome disassembly. In this paper we report cDNA cloning and characterization of mDEAH9, an apparent mammalian homologue of Prp43. Amino acid sequence comparison revealed that the two proteins are ≈65% identical over a 500-aa region spanning the central helicase domain and the C-terminal region. Expression of mDEAH9 in S. cerevisiae bearing a temperature-sensitive mutation in prp43 was sufficient to restore growth at the nonpermissive temperature. This functional complementation was specific, as mouse mDEAH9 failed to complement mutations in related splicing factor genes prp16 or prp22. Finally, double label immunofluorescence experiments performed with mammalian cells revealed colocalization of mDEAH9 and splicing factor SC35 in punctate nuclear speckles. Thus, the hypothesis that mDEAH9 represents the mammalian homologue of yeast Prp43 is supported by its high sequence homology, functional complementation, and colocalization with a known splicing factor in the nucleus. Our results provide additional support for the hypothesis that the spliceosomal machinery that mediates regulated, dynamic changes in conformation of pre-mRNA and snRNP RNAs has been highly conserved through evolution.

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Additional Information:© 1997 The National Academy of Sciences of the USA. Communicated by John N. Abelson, California Institute of Technology, Pasadena, CA, August 8, 1997 (received for review December 17, 1996). This work was supported by National Institutes of Health Grant HL45182 (J.G.C.). The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked ‘‘advertisement’’ in accordance with 18 U.S.C. §1734 solely to indicate this fact.
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Issue or Number:22
PubMed Central ID:PMC23596
Record Number:CaltechAUTHORS:20141125-151704830
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Official Citation:Gee, S., Krauss, S. W., Miller, E., Aoyagi, K., Arenas, J., & Conboy, J. G. (1997). Cloning of mDEAH9, a putative RNA helicase and mammalian homologue of Saccharomyces cerevisiae splicing factor Prp43. Proceedings of the National Academy of Sciences, 94(22), 11803-11807.
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ID Code:52162
Deposited On:26 Nov 2014 15:49
Last Modified:10 Nov 2021 19:21

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