PMID: 9539711May 16, 1998Paper

The human U5-200kD DEXH-box protein unwinds U4/U6 RNA duplices in vitro

Proceedings of the National Academy of Sciences of the United States of America
B LaggerbauerR Lührmann

Abstract

Splicing of nuclear precursors of mRNA (pre-mRNA) involves dynamic interactions between the RNA constituents of the spliceosome. The rearrangement of RNA-RNA interactions, such as the unwinding of the U4/U6 duplex, is believed to be driven by ATP-dependent RNA helicases. We recently have shown that spliceosomal U5 small nuclear ribonucleoproteins (snRNPs) from HeLa cells contain two proteins, U5-200kD and U5-100kD, which share homology with the DEAD/DEXH-box families of RNA helicases. Here we demonstrate that purified U5 snRNPs exhibit ATP-dependent unwinding of U4/U6 RNA duplices in vitro. To identify the protein responsible for this activity, U5 snRNPs were depleted of a subset of proteins under high salt concentrations and assayed for RNA unwinding. The activity was retained in U5 snRNPs that contain the U5-200kD protein but lack U5-100kD, suggesting that the U5-200kD protein could mediate U4/U6 duplex unwinding. Finally, U5-200kD was purified to homogeneity by glycerol gradient centrifugation of U5 snRNP proteins in the presence of sodium thiocyanate, followed by ion exchange chromatography. The RNA unwinding activity was found to reside exclusively with the U5-200kD DEXH-box protein. Our data raise the interesting possibil...Continue Reading

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Citations

Aug 12, 2003·Gene·Edward SilvermanRen-Jang Lin
May 10, 2001·Current Opinion in Cell Biology·C L Will, R Lührmann
Oct 31, 2002·RNA·Rui PengJames G Patton
Apr 22, 2008·Nature Structural & Molecular Biology·Priya BellareErik J Sontheimer
Oct 28, 2008·Nature Structural & Molecular Biology·Irina HäckerReinhard Lührmann
Dec 23, 2008·Nature Structural & Molecular Biology·Corina MaederChristine Guthrie
Jun 16, 2009·Nature Structural & Molecular Biology·Lingdi ZhangRui Zhao
Jan 7, 2009·Nature Structural & Molecular Biology·David A Brow
Apr 28, 2009·Molecular BioSystems·Chaim Wachtel, James L Manley
Oct 10, 2012·Proceedings of the National Academy of Sciences of the United States of America·Karine F SantosMarkus C Wahl
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Oct 11, 2002·The EMBO Journal·Stephanie NottrottReinhard Lührmann
Feb 14, 2002·Nucleic Acids Research·Chun-Hong ChenSoo-Chen Cheng
Aug 22, 2003·Nucleic Acids Research·Stephanie S-I Tseng-RogenskiTien-Hsien Chang
Dec 17, 2005·Genes & Development·Rong-Tzong TsaiSoo-Chen Cheng
Feb 28, 2013·Genes & Development·Gert WeberMarkus C Wahl
Sep 9, 2010·Molecular and Cellular Biology·Yin-Chu ChenMichael C Yu
May 2, 2003·Molecular and Cellular Biology·Hadar MalcaGil Ast
Apr 21, 2005·RNA·Richard J Grainger, Jean D Beggs
Mar 17, 2006·RNA·Tamara J Brenner, Christine Guthrie

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