Binding and cleavage of unstructured RNA by nuclear RNase P
| Autor: | David R. Engelke, Carol A. Fierke, Michael C. Marvin, Scott C. Walker |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Cell Nucleus
RNA Untranslated Base Sequence biology RNase P Nuclear RNase P Molecular Sequence Data Endoribonuclease Saccharomyces cerevisiae Non-coding RNA Molecular biology RNase PH Ribonuclease P Article RNase MRP biology.protein Nucleic Acid Conformation Degradosome RNase H Molecular Biology Protein Binding |
| Zdroj: | RNA. 17:1429-1440 |
| ISSN: | 1469-9001 1355-8382 |
| DOI: | 10.1261/rna.2633611 |
| Popis: | Ribonuclease P (RNase P) is an essential endoribonuclease for which the best-characterized function is processing the 5′ leader of pre-tRNAs. Compared to bacterial RNase P, which contains a single small protein subunit and a large catalytic RNA subunit, eukaryotic nuclear RNase P is more complex, containing nine proteins and an RNA subunit in Saccharomyces cerevisiae. Consistent with this, nuclear RNase P has been shown to possess unique RNA binding capabilities. To understand the unique molecular recognition of nuclear RNase P, the interaction of S. cerevisiae RNase P with single-stranded RNA was characterized. Unstructured, single-stranded RNA inhibits RNase P in a size-dependent manner, suggesting that multiple interactions are required for high affinity binding. Mixed-sequence RNAs from protein-coding regions also bind strongly to the RNase P holoenzyme. However, in contrast to poly(U) homopolymer RNA that is not cleaved, a variety of mixed-sequence RNAs have multiple preferential cleavage sites that do not correspond to identifiable consensus structures or sequences. In addition, pre-tRNATyr, poly(U)50 RNA, and mixed-sequence RNA cross-link with purified RNase P in the RNA subunit Rpr1 near the active site in “Conserved Region I,” although the exact positions vary. Additional contacts between poly(U)50 and the RNase P proteins Rpr2p and Pop4p were identified. We conclude that unstructured RNAs interact with multiple protein and RNA contacts near the RNase P RNA active site, but that cleavage depends on the nature of interaction with the active site. |
| Databáze: | OpenAIRE |
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