PMID: 8600454Mar 1, 1996Paper

Identification of an RNA-dependent ATPase activity in mammalian U5 snRNPs

Nucleic Acids Research
B LaggerbauerR Lührmann

Abstract

Nuclear pre-mRNA splicing requires ATP at several steps from spliceosome assembly to product release. Here, we demonstrate that an integral component of the 20S U5 snRNP is an RNA-dependent ATPase. The ATPase activity of 20S U5 and 25S [U4/U6.U5] snRNPs purified by glycerol gradient centrifugation is strongly stimulated by homopolymeric RNA but not ssDNA. Purified 12S Ul and U2 snRNPs do not exhibit ATPase activity. Moreover, the U5-associated NTPase specifically hydrolyzes ATP and dATP. The additional purification of 20S U5 snRNPs by Mono Q chromatography does not affect the efficiency of ATP hydrolysis. Both U5 and tri-snRNPs bind ATP stoichiometrically in an RNA-independent manner. A candidate ATPase was identified by UV-irradiation of purified snRNPs with radiolabeled ATP. In the presence of homopolymeric RNA, the 200 kDa U5-specific protein is the major crosslinked protein, even in Mono Q-purified U5 snRNPs. The correlation between RNA-dependent ATPase activity in the U5 snRNP and the RNA-dependent onset of this crosslink strongly suggests that the 200 kDa protein is an RNA-dependent ATPase. Furthermore, both the formation of the crosslink and ATPase activity appear with a similar substrate specificity for ATP.

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Citations

Jun 1, 1997·Current Opinion in Cell Biology·C L Will, R Lührmann
Jun 10, 2010·The Journal of Biological Chemistry·Christian Matranga, Anna Marie Pyle
Oct 6, 1997·The EMBO Journal·A J Newman
Apr 25, 2002·Molecular and Cellular Biology·Claudia SchneiderReinhard Lührmann
Oct 29, 1997·Proceedings of the National Academy of Sciences of the United States of America·S GeeJ G Conboy
Jun 11, 2014·Acta Crystallographica. Section D, Biological Crystallography·Sina MöhlmannRalf Ficner
May 16, 1998·Proceedings of the National Academy of Sciences of the United States of America·B LaggerbauerR Lührmann
Jun 22, 1999·Journal of Molecular Biology·R L Gonzalez, I Tinoco
Mar 3, 2004·Nature Reviews. Molecular Cell Biology·Sanda Rocak, Patrick Linder
May 9, 2003·The Journal of Biological Chemistry·Cornelia BartelsPatrizia Fabrizio
Jun 1, 1997·Molecular and Cellular Biology·J VenemaD Tollervey

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