Nucleic Acids Research, Vol 24, Issue 5 868-875, Copyright © 1996 by Oxford University Press
B Laggerbauer, J Lauber and R Luhrmann
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.
ARTICLES
Identification of an RNA-dependent ATPase activity in mammalian U5 snRNPs
Institut fur Molekularbiologie und Tumorforschung, Marburg, Germany.
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