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RNA11 protein is associated with the yeast spliceosome and is localized in the periphery of the cell nucleus

Chang, Tien-Hsien and Clark, Michael W. and Lustig, Arthur J. and Cusick, Michael E. and Abelson, John (1988) RNA11 protein is associated with the yeast spliceosome and is localized in the periphery of the cell nucleus. Molecular and Cellular Biology, 8 (6). pp. 2379-2393. ISSN 0270-7306.

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The yeast rna mutations (rna2 through rna10/11) are a set of temperature-sensitive mutations that result in the accumulation of pre-mRNAs at the nonpermissive temperature. Most of the yeast RNA gene products are involved in and essential for mRNA splicing in vitro, suggesting that they code for components of the splicing machinery. We tested this proposal by using an in vitro-synthesized RNA11 protein to complement the temperature-sensitive defect of the rna11 extract. During the in vitro complementation, the input RNA11 protein was associated with the 40S spliceosome and a 30S complex, suggesting that the RNA11 protein is indeed a component of the spliceosome. The formation of the RNA11-associated 30S complex did not require any exogenous RNA substrate, suggesting that this 30S particle is likely to be a preassembled complex involved in splicing. The RNA11-specific antibody inhibited the mRNA splicing in vitro, confirming the essential role of the RNA11 protein in mRNA splicing. Finally, using the anti-RNA11 antibody, we localized the RNA11 protein to the periphery of the yeast nucleus.

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Additional Information:Copyright © 1988 by the American Society for Microbiology. Received 7 January 1988/Accepted 9 March 1988 We thank R. Luhrmann and S. Hoch for providing anti-m3G and anti-Sm (no. 58) antisera, respectively, and R. Sternglanz for supplying the cloned RNAII gene. We also thank E. Grayhack, S. Goelz, S.-C. Cheng, R.-J. Lin, J. Dahlberg, and S. Westaway for their stimulating discussions and technical advice. The critical reading of the manuscript by U. Vijayraghavan and K. Tanner is also greatly appreciated. We thank D. McPheeters for pointing out the potential zinc finger motif in the RNA11 protein. Computer resources were provided by BIONET National Computer Resource for Molecular Biology, whose funding is provided by the Biomedical Research Technology Program, Division of Research Resources, National Institutes of Health (grant 1-U41RR-01685-05). M.E.C. is supported by a postdoctoral fellowship (S-12-86) from the American Cancer Society. This work was supported entirely by a Public Health Service grant awarded to J.A. from the National Institutes of Health.
Issue or Number:6
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ID Code:2520
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Deposited On:06 Apr 2006
Last Modified:02 Oct 2019 22:54

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