Assembly of Mitochondrial Ribonucleoprotein Complexes Involves Specific Guide RNA (gRNA)-Binding Proteins and gRNA Domains but Does Not Require Preedited mRNA
| Autor: | H U Göringer, Kenneth Stuart, Laurie K. Read |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Genetics
Messenger RNA Genes Protozoan Trypanosoma brucei brucei RNA-Binding Proteins RNA-binding protein Cell Biology Mitochondrion Biology Blotting Northern DNA-binding protein In vitro Mitochondria Cell biology Ribonucleoproteins RNA editing Animals RNA Messenger Guide RNA Molecular Biology Gene Deletion RNA Protozoan RNA Guide Kinetoplastida Research Article Ribonucleoprotein |
| Zdroj: | Molecular and Cellular Biology. 14:2629-2639 |
| ISSN: | 1098-5549 |
| DOI: | 10.1128/mcb.14.4.2629-2639.1994 |
| Popis: | RNA editing in kinetoplastids probably employs a macromolecular complex, the editosome, that is likely to include the guide RNAs (gRNAs) which specify the edited sequence. Specific ribonucleoprotein (RNP) complexes which form in vitro with gRNAs (H. U. Göringer, D. J. Koslowsky, T. H. Morales, and K. D. Stuart, Proc. Natl. Acad. Sci. USA, in press) are potential editosomes or their precursors. We find that several factors are important for in vitro formation of these RNP complexes and identify specific gRNA-binding proteins present in the complexes. Preedited mRNA promotes the in vitro formation of the four major gRNA-containing RNP complexes under some conditions but is required for the formation of only a subcomponent of one complex. The 5' gRNA sequence encompassing the RYAYA and anchor regions and the 3' gRNA oligo(U) tail are both important in complex formation, since their deletion results in a dramatic decrease of some complexes and the absence of others. UV cross-linking experiments identify several proteins which are in contact with gRNA and preedited mRNA in mitochondrial extracts. Proteins of 25 and 90 kDa are highly specific for gRNAs, and the 90-kDa protein binds specifically to gRNA oligo(U) tails. The gRNA-binding proteins exhibit a differential distribution between the four in vitro-formed complexes. These experiments reveal several proteins potentially involved in RNA editing and indicate that multiple recognition elements in gRNAs are used for complex formation. |
| Databáze: | OpenAIRE |
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