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snRNA interactions at 5' and 3' splice sites monitored by photoactivated crosslinking in yeast spliceosomes.
RNA. 1995 Nov; 1(9):968-80.RNA

Abstract

Splice site recognition and catalysis of the transesterification reactions in the spliceosome are accompanied by a dynamic series of interactions involving conserved or invariant sequences in the spliceosomal snRNAs. We have used site-specific photoactivated crosslinking in yeast spliceosomes to monitor interactions between snRNAs and exon sequences near the 5' and 3' splice sites. The last nucleotide of the 5' exon can be crosslinked to an invariant loop sequence in U5 SnRNA before and after 5' splice site cleavage. The first nucleotide of the 3' exon can also be crosslinked to the same U5 loop sequence, but this contact is only detectable after the first transesterification. These results are in close agreement with earlier data from mammalian splicing extracts, and they are consistent with a model in which U5 snRNA aligns the 5' and 3' exons for the second transesterification. After the first catalytic step of splicing, the first nucleotide of the 3' exon can also crosslink to nt U23 in U2 snRNA. This is one of a cluster of residues in U2-U6 helix I implicated by mutational analysis in the second catalytic step of splicing. The crosslinking data suggest that these residues in U2-U6 helix I are in close proximity to the scissile phosphodiester bond at the 3' splice site prior to the second transesterification. These results constitute the first biochemical evidence for a direct interaction between the 3' splice site and U2 snRNA.

Authors+Show Affiliations

MRC Laboratory of Molecular Biology, Cambridge, United Kingdom.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

8548661

Citation

Newman, A J., et al. "SnRNA Interactions at 5' and 3' Splice Sites Monitored By Photoactivated Crosslinking in Yeast Spliceosomes." RNA (New York, N.Y.), vol. 1, no. 9, 1995, pp. 968-80.
Newman AJ, Teigelkamp S, Beggs JD. SnRNA interactions at 5' and 3' splice sites monitored by photoactivated crosslinking in yeast spliceosomes. RNA. 1995;1(9):968-80.
Newman, A. J., Teigelkamp, S., & Beggs, J. D. (1995). SnRNA interactions at 5' and 3' splice sites monitored by photoactivated crosslinking in yeast spliceosomes. RNA (New York, N.Y.), 1(9), 968-80.
Newman AJ, Teigelkamp S, Beggs JD. SnRNA Interactions at 5' and 3' Splice Sites Monitored By Photoactivated Crosslinking in Yeast Spliceosomes. RNA. 1995;1(9):968-80. PubMed PMID: 8548661.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - snRNA interactions at 5' and 3' splice sites monitored by photoactivated crosslinking in yeast spliceosomes. AU - Newman,A J, AU - Teigelkamp,S, AU - Beggs,J D, PY - 1995/11/1/pubmed PY - 1995/11/1/medline PY - 1995/11/1/entrez SP - 968 EP - 80 JF - RNA (New York, N.Y.) JO - RNA VL - 1 IS - 9 N2 - Splice site recognition and catalysis of the transesterification reactions in the spliceosome are accompanied by a dynamic series of interactions involving conserved or invariant sequences in the spliceosomal snRNAs. We have used site-specific photoactivated crosslinking in yeast spliceosomes to monitor interactions between snRNAs and exon sequences near the 5' and 3' splice sites. The last nucleotide of the 5' exon can be crosslinked to an invariant loop sequence in U5 SnRNA before and after 5' splice site cleavage. The first nucleotide of the 3' exon can also be crosslinked to the same U5 loop sequence, but this contact is only detectable after the first transesterification. These results are in close agreement with earlier data from mammalian splicing extracts, and they are consistent with a model in which U5 snRNA aligns the 5' and 3' exons for the second transesterification. After the first catalytic step of splicing, the first nucleotide of the 3' exon can also crosslink to nt U23 in U2 snRNA. This is one of a cluster of residues in U2-U6 helix I implicated by mutational analysis in the second catalytic step of splicing. The crosslinking data suggest that these residues in U2-U6 helix I are in close proximity to the scissile phosphodiester bond at the 3' splice site prior to the second transesterification. These results constitute the first biochemical evidence for a direct interaction between the 3' splice site and U2 snRNA. SN - 1355-8382 UR - https://www.unboundmedicine.com/medline/citation/8548661/snRNA_interactions_at_5'_and_3'_splice_sites_monitored_by_photoactivated_crosslinking_in_yeast_spliceosomes_ L2 - http://www.rnajournal.org/cgi/pmidlookup?view=long&pmid=8548661 DB - PRIME DP - Unbound Medicine ER -