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Two Domains of Yeast U6 Small Nuclear RNA Required for Both Steps of Nuclear Precursor Messenger RNA Splicing

Fabrizio, Patrizia and Abelson, John (1990) Two Domains of Yeast U6 Small Nuclear RNA Required for Both Steps of Nuclear Precursor Messenger RNA Splicing. Science, 250 (4979). pp. 404-409. ISSN 0036-8075. doi:10.1126/science.2145630.

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U6 is one of the five small nuclear RNA's (snRNA's) that are required for splicing of nuclear precursor messenger RNA (pre-mRNA). The size and sequence of U6 RNA are conserved among organisms as diverse as yeast and man, and so it has been proposed that U6 RNA functions as a catalytic element in splicing. A procedure for in vitro reconstitution of functional yeast U6 small nuclear ribonucleoproteins (snRNP's) with synthetic U6 RNA was applied in an attempt to elucidate the function of yeast U6 RNA. Two domains in U6 RNA were identified, each of which is required for in vitro splicing. Single nucleotide substitutions in these two domains block splicing either at the first or the second step. Invariably, U6 RNA mutants that block the first step of splicing do not enter the spliceosome. On the other hand, those that block the second step of splicing form a spliceosome but block cleavage at the 3' splice site of the intron. In both domains, the positions of base changes that block the second step of splicing correspond exactly to the site of insertion of pre-mRNA-type introns into the U6 gene of two yeast species, providing a possible explanation for the mechanism of how these introns originated and adding further evidence for the proposed catalytic role of U6 RNA.

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Additional Information:© 1990 American Association for the Advancement of Science. Received 16 May 1990; accepted 7 September 1990. We thank R. Rauhut, H. Madhani, R. Bordonne, D. Horowitz, C. O'Day, G. Dalbadie-McFarland, J. Arenas, L. Krinke, S. Ruby, and D. McPheeters for comments on the manuscript; R. Bordonne, H. Madhani, and C. Guthrie for suggestions and for discussing unpublished results; I. Mattaj and A. Bindereif for sharing unpublished results; and S. Horvath and staff for synthesis of oligonucleotides. Supported by NIH grant GM 32637 to J.A.
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NIHGM 32637
Issue or Number:4979
Record Number:CaltechAUTHORS:20141205-133513787
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Official Citation:Two domains of yeast U6 small nuclear RNA required for both steps of nuclear precursor messenger RNA splicing P Fabrizio and J Abelson Science 19 October 1990: 250 (4979), 404-409. [DOI:10.1126/science.2145630]
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:52439
Deposited By: Tony Diaz
Deposited On:09 Dec 2014 22:41
Last Modified:10 Nov 2021 19:41

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