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Extensive interactions of PRP8 protein with the 5' and 3' splice sites during splicing suggest a role in stabilization of exon alignment by U5 snRNA.
EMBO J. 1995 Jun 01; 14(11):2602-12.EJ

Abstract

Precursor RNAs containing 4-thiouridine at specific sites were used with UV-crosslinking to map the binding sites of the yeast protein splicing factor PRP8. PRP8 protein interacts with a region of at least eight exon nucleotides at the 5' splice site and a minimum of 13 exon nucleotides and part of the polypyrimidine tract in the 3' splice site region. Crosslinking of PRP8 to mutant and duplicated 3' splice sites indicated that the interaction is not sequence specific, nor does it depend on the splice site being functional. Binding of PRP8 to the 5' exon was established before step 1 and to the 3' splice site region after step 1 of splicing. These interactions place PRP8 close to the proposed catalytic core of the spliceosome during both transesterification reactions. To date, this represents the most extensive mapping of the binding site(s) of a splicing factor on the substrate RNA. We propose that the large binding sites of PRP8 stabilize the intrinsically weaker interactions of U5 snRNA with both exons at the splice sites for exon alignment by the U5 snRNP.

Authors+Show Affiliations

Institute of Cell and Molecular Biology, University of Edinburgh, Cambridge, UK.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

7781612

Citation

Teigelkamp, S, et al. "Extensive Interactions of PRP8 Protein With the 5' and 3' Splice Sites During Splicing Suggest a Role in Stabilization of Exon Alignment By U5 SnRNA." The EMBO Journal, vol. 14, no. 11, 1995, pp. 2602-12.
Teigelkamp S, Newman AJ, Beggs JD. Extensive interactions of PRP8 protein with the 5' and 3' splice sites during splicing suggest a role in stabilization of exon alignment by U5 snRNA. EMBO J. 1995;14(11):2602-12.
Teigelkamp, S., Newman, A. J., & Beggs, J. D. (1995). Extensive interactions of PRP8 protein with the 5' and 3' splice sites during splicing suggest a role in stabilization of exon alignment by U5 snRNA. The EMBO Journal, 14(11), 2602-12.
Teigelkamp S, Newman AJ, Beggs JD. Extensive Interactions of PRP8 Protein With the 5' and 3' Splice Sites During Splicing Suggest a Role in Stabilization of Exon Alignment By U5 SnRNA. EMBO J. 1995 Jun 1;14(11):2602-12. PubMed PMID: 7781612.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Extensive interactions of PRP8 protein with the 5' and 3' splice sites during splicing suggest a role in stabilization of exon alignment by U5 snRNA. AU - Teigelkamp,S, AU - Newman,A J, AU - Beggs,J D, PY - 1995/6/1/pubmed PY - 1995/6/1/medline PY - 1995/6/1/entrez SP - 2602 EP - 12 JF - The EMBO journal JO - EMBO J VL - 14 IS - 11 N2 - Precursor RNAs containing 4-thiouridine at specific sites were used with UV-crosslinking to map the binding sites of the yeast protein splicing factor PRP8. PRP8 protein interacts with a region of at least eight exon nucleotides at the 5' splice site and a minimum of 13 exon nucleotides and part of the polypyrimidine tract in the 3' splice site region. Crosslinking of PRP8 to mutant and duplicated 3' splice sites indicated that the interaction is not sequence specific, nor does it depend on the splice site being functional. Binding of PRP8 to the 5' exon was established before step 1 and to the 3' splice site region after step 1 of splicing. These interactions place PRP8 close to the proposed catalytic core of the spliceosome during both transesterification reactions. To date, this represents the most extensive mapping of the binding site(s) of a splicing factor on the substrate RNA. We propose that the large binding sites of PRP8 stabilize the intrinsically weaker interactions of U5 snRNA with both exons at the splice sites for exon alignment by the U5 snRNP. SN - 0261-4189 UR - https://www.unboundmedicine.com/medline/citation/7781612/Extensive_interactions_of_PRP8_protein_with_the_5'_and_3'_splice_sites_during_splicing_suggest_a_role_in_stabilization_of_exon_alignment_by_U5_snRNA_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0261-4189&date=1995&volume=14&issue=11&spage=2602 DB - PRIME DP - Unbound Medicine ER -