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Prp21, a U2-snRNP-associated protein, and Prp24, a U6-snRNP-associated protein, functionally interact during spliceosome assembly in yeast

Vaijayanti, Vaidya C and Usha, Vijayraghavan (1998) Prp21, a U2-snRNP-associated protein, and Prp24, a U6-snRNP-associated protein, functionally interact during spliceosome assembly in yeast. In: Journal of Genetics, 77 (2-3). pp. 85-94.

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Abstract

Earlier studies on genetic suppression of prp24-1 by prp21-2 suggested an association between yeast Prp21 and Prp24 proteins, which are associated, respectively, with U2 snRNA and U6 snRNA. Here we report analyses of physical and functional interaction between these factors. Missense mutations in functionally important domains reside in prp21-2 and prp24-1. Two-hybrid assays do not detect interaction between wild-type or mutant proteins. Prp21-2 and Prp24-1 protein in prp21-2 or prp24-1 extracts can be heat-inactivated in vitro. In contrast, heat-treated extracts from the revertant strain prp21-2 prp24-1 demonstrate allele-specific restoration of splicing. Suppression of prp24-1 by prp21-2 does not cause coimmunoprecipitation of U2 and U6 snRNAs. We demonstrate the presence of Prp21 in the spliceosome assembly intermediate A2-1, and our data suggest the presence of Prp24 in the same complex. Kinetic analysis of assembly in heat-treated revertant extracts reveal a rate-limiting conversion of complex B to A2-1, suggesting transient association between the mutant proteins at this step. Our data also imply a requirement for Prp21 during B to A2-1 conversion. We conclude that a transient yet likely functional association between Prp21 and Prp24 occurs during spliceosome assembly.

Item Type: Journal Article
Publication: Journal of Genetics
Publisher: Indian Academy of Sciences
Additional Information: copyright of thia article belongs to Indian Academy of Sciences.
Keywords: yeast;pre-mRNA splicing;PRP21;PRP24;protein interactions.
Department/Centre: Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited: 11 Oct 2007
Last Modified: 07 Feb 2012 06:34
URI: http://eprints.iisc.ac.in/id/eprint/12211

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