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Analysis of Snu13p mutations reveals differential interactions with the U4 snRNA and U3 snoRNA.
RNA. 2004 Feb; 10(2):308-20.RNA

Abstract

Pre-mRNA splicing is executed by the spliceosome, a complex of small nuclear RNAs (snRNAs) and numerous proteins. One such protein, 15.5K/Snu13p, is associated with the spliceosomal U4/U6.U5 tri-snRNP and box C/D small nucleolar ribonucleoprotein particles (snoRNPs), which act during preribosomal RNA (rRNA) processing. As such, it is the first splicing factor to be identified in two functionally distinct particles. 15.5K binds to an internal helix-bulge-helix (K-turn) structure in the U4 snRNA and two such structures in the U3 snoRNA. Previous work has concentrated on the structural basis of the interaction of 15.5K with the RNAs and has been carried out in vitro. Here we present a functional analysis of Snu13p in vivo, using a galactose inducible SNU13 strain to investigate the basis of three lethal mutations in Saccharomyces cerevisiae. Two are point mutations that map to the RNA-binding domain, and the third is a C-terminal deletion. These mutations result in accumulation of unspliced pre-mRNA, confirming a role for Snu13p in pre-mRNA splicing. In addition, these mutants also display rRNA processing defects that are variable in nature. Analysis of one mutant in the RNA-binding domain reveals a reduction in the levels of the U4 snRNA, U6 snRNA, and box C/D snoRNAs, but not H/ACA snoRNAs, supporting a role for Snu13p in accumulation and/or maintenance of specific RNAs. The mutations in the RNA-binding domain exhibit differential binding to the U4 snRNA and U3 snoRNA in vitro, suggesting that there are differences in the mode of interaction of Snu13p with these two RNAs.

Authors+Show Affiliations

School of Biological Sciences, University of Manchester, Manchester, M13 9PT, UK.No affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

14730029

Citation

Dobbyn, Helen C., and Raymond T. O'Keefe. "Analysis of Snu13p Mutations Reveals Differential Interactions With the U4 snRNA and U3 SnoRNA." RNA (New York, N.Y.), vol. 10, no. 2, 2004, pp. 308-20.
Dobbyn HC, O'Keefe RT. Analysis of Snu13p mutations reveals differential interactions with the U4 snRNA and U3 snoRNA. RNA. 2004;10(2):308-20.
Dobbyn, H. C., & O'Keefe, R. T. (2004). Analysis of Snu13p mutations reveals differential interactions with the U4 snRNA and U3 snoRNA. RNA (New York, N.Y.), 10(2), 308-20.
Dobbyn HC, O'Keefe RT. Analysis of Snu13p Mutations Reveals Differential Interactions With the U4 snRNA and U3 SnoRNA. RNA. 2004;10(2):308-20. PubMed PMID: 14730029.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Analysis of Snu13p mutations reveals differential interactions with the U4 snRNA and U3 snoRNA. AU - Dobbyn,Helen C, AU - O'Keefe,Raymond T, PY - 2004/1/20/pubmed PY - 2004/2/26/medline PY - 2004/1/20/entrez SP - 308 EP - 20 JF - RNA (New York, N.Y.) JO - RNA VL - 10 IS - 2 N2 - Pre-mRNA splicing is executed by the spliceosome, a complex of small nuclear RNAs (snRNAs) and numerous proteins. One such protein, 15.5K/Snu13p, is associated with the spliceosomal U4/U6.U5 tri-snRNP and box C/D small nucleolar ribonucleoprotein particles (snoRNPs), which act during preribosomal RNA (rRNA) processing. As such, it is the first splicing factor to be identified in two functionally distinct particles. 15.5K binds to an internal helix-bulge-helix (K-turn) structure in the U4 snRNA and two such structures in the U3 snoRNA. Previous work has concentrated on the structural basis of the interaction of 15.5K with the RNAs and has been carried out in vitro. Here we present a functional analysis of Snu13p in vivo, using a galactose inducible SNU13 strain to investigate the basis of three lethal mutations in Saccharomyces cerevisiae. Two are point mutations that map to the RNA-binding domain, and the third is a C-terminal deletion. These mutations result in accumulation of unspliced pre-mRNA, confirming a role for Snu13p in pre-mRNA splicing. In addition, these mutants also display rRNA processing defects that are variable in nature. Analysis of one mutant in the RNA-binding domain reveals a reduction in the levels of the U4 snRNA, U6 snRNA, and box C/D snoRNAs, but not H/ACA snoRNAs, supporting a role for Snu13p in accumulation and/or maintenance of specific RNAs. The mutations in the RNA-binding domain exhibit differential binding to the U4 snRNA and U3 snoRNA in vitro, suggesting that there are differences in the mode of interaction of Snu13p with these two RNAs. SN - 1355-8382 UR - https://www.unboundmedicine.com/medline/citation/14730029/Analysis_of_Snu13p_mutations_reveals_differential_interactions_with_the_U4_snRNA_and_U3_snoRNA_ DB - PRIME DP - Unbound Medicine ER -