Reference: Karaduman R, et al. (2006) RNA structure and RNA-protein interactions in purified yeast U6 snRNPs. J Mol Biol 356(5):1248-62

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Abstract


The U6 small nuclear RNA (snRNA) undergoes major conformational changes during the assembly of the spliceosome and catalysis of splicing. It associates with the specific protein Prp24p, and a set of seven LSm2p-8p proteins, to form the U6 small nuclear ribonucleoprotein (snRNP). These proteins have been proposed to act as RNA chaperones that stimulate pairing of U6 with U4 snRNA to form the intermolecular stem I and stem II of the U4/U6 duplex, whose formation is essential for spliceosomal function. However, the mechanism whereby Prp24p and the LSm complex facilitate U4/U6 base-pairing, as well as the exact binding site(s) of Prp24p in the native U6 snRNP, are not well understood. Here, we have investigated the secondary structure of the U6 snRNA in purified U6 snRNPs and compared it with its naked form. Using RNA structure-probing techniques, we demonstrate that within the U6 snRNP a large internal region of the U6 snRNA is unpaired and protected from chemical modification by bound Prp24p. Several of these U6 nucleotides are available for base-pairing interaction, as only their sugar backbone is contacted by Prp24p. Thus, Prp24p can present them to the U4 snRNA and facilitate formation of U4/U6 stem I. We show that the 3' stem-loop is not bound strongly by U6 proteins in native particles. However, when compared to the 3' stem-loop in the naked U6 snRNA, it has a more open conformation, which would facilitate formation of stem II with the U4 snRNA. Our data suggest that the combined association of Prp24p and the LSm complex confers upon U6 nucleotides a conformation favourable for U4/U6 base-pairing. Interestingly, we find that the open structure of the yeast U6 snRNA in native snRNPs can also be adopted by human U6 and U6atac snRNAs.

Reference Type
Journal Article | Research Support, Non-U.S. Gov't
Authors
Karaduman R, Fabrizio P, Hartmuth K, Urlaub H, Lührmann R
Primary Lit For
PRP24 | U6 small nuclear ribonucleoprotein complex | U4/U6 small nuclear ribonucleoprotein complex | Sm complex
Additional Lit For
SNR6 | LSM8 | LSM2 | LSM4 | LSM5 | LSM7 | LSM3

Gene Ontology Annotations 1 entry for 1 gene


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Gene/ComplexQualifierGene Ontology TermAnnotation ExtensionEvidenceSourceAssigned On
PRP24enablessnRNA bindinghas input SNR6IPI with SNR6SGD2013-08-07
Showing 1 to 1 of 1 entries

Interaction Annotations


Genetic Interactions

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Interactor Interactor Allele Assay Annotation Action Phenotype SGA score P-value Source Reference

Physical Interactions 7 entries for 8 genes

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InteractorInteractorAssayAnnotationActionModification
LSM2PRP24Affinity Capture-RNAmanually curatedHit-BaitNo Modification
LSM4PRP24Affinity Capture-RNAmanually curatedHit-BaitNo Modification
LSM5PRP24Affinity Capture-RNAmanually curatedHit-BaitNo Modification
LSM6PRP24Affinity Capture-RNAmanually curatedHit-BaitNo Modification
LSM8PRP24Affinity Capture-RNAmanually curatedHit-BaitNo Modification
PRP24LSM3Affinity Capture-RNAmanually curatedBait-HitNo Modification
PRP24LSM7Affinity Capture-RNAmanually curatedBait-HitNo Modification
Showing 1 to 7 of 7 entries