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Analysis of Small Nuclear RNAs in a Precatalytic Spliceosome

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U1 small nuclear RNA plays an important role in early stages of intron recognition and spliceosome assembly. The 5 splice site of the premessenger RNA base-pairs with the 5 end of Ul; however, that interaction appears to be replaced by U5 and U6 at later stages of the splicing process. It has not been established when this transition occurs nor what factors are required for the transition. The PRP2 gene of Saccharomyces cerevisiae encodes an RNA-dependent ATPase that belongs to the DEAH putative RNA helicase family. A spliceosome can be assembled in the absence of PRP2 but the ATPase activity of PRP2 is required for the onset of the catalytic reactions in the spliceosome. By probing the precatalytic spliceosome formed in temperature-sensitive PRP2 mutant extracts with oligonucleotides complementary to snRNAs, we found that the 5 end of Ul was sensitive to RNase H digestion whereas the 5 splice site-interacting region of U6 became resistant. Furthermore, by treating with heparin, a spliceosome depleted of Ul snRNA was isolated that subsequently underwent splicing with additional protein factors and ATP. Thus, these results indicate that PRP2 is not responsible for the transition from Ul to U6 in the spliceosome and that the entire Ul snRNA is not involved in the catalytic phase of pre-mRNA splicing.
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Keywords: DEAH proteins; PRP2 ATPase; Pre-mRNA splicing Saccharomyces cerevisiae; Small nuclear RNAs; Spliceosome

Document Type: Research Article

Publication date: January 1, 1996

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  • Gene Expression, The Journal of Liver Research will publish articles in all aspects of hepatology. Hepatology, as a research discipline, has seen unprecedented growth especially in the cellular and molecular mechanisms of hepatic health and disease, which continues to have a major impact on understanding liver development, stem cells, carcinogenesis, tissue engineering, injury, repair, regeneration, immunology, metabolism, fibrosis, and transplantation. Continued research and improved understanding in these areas will have a meaningful impact on liver disease prevention, diagnosis, and treatment. The existing journal Gene Expression has expanded its focus to become Gene Expression, The Journal of Liver Research to meet this growing demand. In its revised and expanded scope, the journal will publish high-impact original articles, reviews, short but complete articles, and special articles (editorials, commentaries, opinions) on all aspects of hepatology, making it a unique and invaluable resource for readers interested in this field. The expanded team, led by an Editor-in-Chief who is uniquely qualified and a renowned expert, along with a dynamic and functional editorial board, is determined to make this a premier journal in the field of hepatology.
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