Genome-wide search for yeast RNase P substrates reveals role in maturation of intron-encoded box C/D small nucleolar RNAs
| Autor: | Gregg B. Whitworth, Jeffrey A. Pleiss, David R. Engelke, Daniel J. Coughlin, Scott C. Walker |
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| Rok vydání: | 2008 |
| Předmět: |
Multidisciplinary
biology Exosome complex Nuclear RNase P RNase P RNA Splicing Saccharomyces cerevisiae Biological Sciences Non-coding RNA RNase PH Molecular biology Ribonuclease P RNase MRP Biochemistry Mutation biology.protein RNA Small Nucleolar Genome Fungal Small nucleolar RNA RNase H Oligonucleotide Array Sequence Analysis |
| Zdroj: | Proceedings of the National Academy of Sciences. 105:12218-12223 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.0801906105 |
| Popis: | Ribonuclease P (RNase P) is an essential endonuclease responsible for the 5′-end maturation of precursor tRNAs. Bacterial RNase P also processes precursor 4.5S RNA, tmRNA, 30S preribosomal RNA, and several reported protein-coding RNAs. Eukaryotic nuclear RNase P is far more complex than in the bacterial form, employing multiple essential protein subunits in addition to the catalytic RNA subunit. RNomic studies have shown that RNase P binds other RNAs in addition to tRNAs, but no non-tRNA substrates have previously been identified. Additional substrates were identified by using a multipronged approach in the budding yeast Saccharomyces cerevisiae . First, RNase P-dependant changes in RNA abundance were examined on whole-genome microarrays by using strains containing temperature sensitive (TS) mutations in two of the essential RNase P subunits, Pop1p and Rpr1r. Second, RNase P was rapidly affinity-purified, and copurified RNAs were identified by using a genome-wide microarray. Third, to identify RNAs that do not change abundance when RNase P is depleted but accumulate as larger precursors, >80 potential small RNA substrates were probed directly by Northern blot analysis with RNA from the RNase P TS mutants. Numerous potential substrates were identified, of which we characterized the box C/D intron-encoded small nucleolar RNAs (snoRNAs), because these both copurify with RNase P and accumulate larger forms in the RNase P temperature-sensitive mutants. It was previously known that two pathways existed for excising these snoRNAs, one using the pre-mRNA splicing path and the other that was independent of splicing. RNase P appears to participate in the splicing-independent path for the box C/D intron-encoded snoRNAs. |
| Databáze: | OpenAIRE |
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