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Interaction of the yeast DExH-box RNA helicase prp22p with the 3' splice site during the second step of nuclear pre-mRNA splicing.
Nucleic Acids Res. 2000 Mar 15; 28(6):1313-21.NA

Abstract

Using site-specific incorporation of the photo-chemical cross-linking reagent 4-thiouridine, we demonstrate the previously unknown association of two proteins with yeast 3' splice sites. One of these is an unidentified approximately 122 kDa protein that cross-links to 3' splice sites during formation of the pre--spliceosome. The other factor is the DExH-box RNA helicase, Prp22p. With substrates functional in the second step of splicing, only very weak cross-linking of Prp22p to intron sequences at the 3' splice site is observed. In contrast, substrates blocked at the second step exhibit strong cross-linking of Prp22 to intron sequences at the 3' splice site, but not to adjacent exon sequences. In vitro reconstitution experiments also show that the association of Prp22p with intron sequences at the 3' splice site is dependent on Prp16p and does not persist when release of mature mRNA from the spliceosome is blocked. Taken together, these results suggest that the 3' splice site of yeast introns is contacted much earlier than previously envisioned by a protein of approximately 120 kDa, and that a transient association of Prp22p with the 3' splice site occurs between the first and second catalytic steps.

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Authors+Show Affiliations

McPheeters DS
Department of Biochemistry and the Center for RNA Moelcular Biology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA. dsm10@po.cwru.edu
Schwer B
No affiliation info available
Muhlenkamp P
No affiliation info available

MeSH

ActinsAdenosine TriphosphatasesAdenosine TriphosphateBase SequenceCatalysisCell NucleusCross-Linking ReagentsDEAD-box RNA HelicasesExonsFungal ProteinsIntronsMolecular WeightMutationRNA HelicasesRNA PrecursorsRNA SplicingRNA Splicing FactorsRNA, FungalRNA, MessengerRNA-Binding ProteinsRegulatory Sequences, Nucleic AcidSaccharomyces cerevisiaeSaccharomyces cerevisiae ProteinsSpliceosomesThiouridine

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

10684925

Citation

McPheeters, D S., et al. "Interaction of the Yeast DExH-box RNA Helicase Prp22p With the 3' Splice Site During the Second Step of Nuclear pre-mRNA Splicing." Nucleic Acids Research, vol. 28, no. 6, 2000, pp. 1313-21.
McPheeters DS, Schwer B, Muhlenkamp P. Interaction of the yeast DExH-box RNA helicase prp22p with the 3' splice site during the second step of nuclear pre-mRNA splicing. Nucleic Acids Res. 2000;28(6):1313-21.
McPheeters, D. S., Schwer, B., & Muhlenkamp, P. (2000). Interaction of the yeast DExH-box RNA helicase prp22p with the 3' splice site during the second step of nuclear pre-mRNA splicing. Nucleic Acids Research, 28(6), 1313-21.
McPheeters DS, Schwer B, Muhlenkamp P. Interaction of the Yeast DExH-box RNA Helicase Prp22p With the 3' Splice Site During the Second Step of Nuclear pre-mRNA Splicing. Nucleic Acids Res. 2000 Mar 15;28(6):1313-21. PubMed PMID: 10684925.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Interaction of the yeast DExH-box RNA helicase prp22p with the 3' splice site during the second step of nuclear pre-mRNA splicing. AU - McPheeters,D S, AU - Schwer,B, AU - Muhlenkamp,P, PY - 2000/2/24/pubmed PY - 2000/4/25/medline PY - 2000/2/24/entrez SP - 1313 EP - 21 JF - Nucleic acids research JO - Nucleic Acids Res VL - 28 IS - 6 N2 - Using site-specific incorporation of the photo-chemical cross-linking reagent 4-thiouridine, we demonstrate the previously unknown association of two proteins with yeast 3' splice sites. One of these is an unidentified approximately 122 kDa protein that cross-links to 3' splice sites during formation of the pre--spliceosome. The other factor is the DExH-box RNA helicase, Prp22p. With substrates functional in the second step of splicing, only very weak cross-linking of Prp22p to intron sequences at the 3' splice site is observed. In contrast, substrates blocked at the second step exhibit strong cross-linking of Prp22 to intron sequences at the 3' splice site, but not to adjacent exon sequences. In vitro reconstitution experiments also show that the association of Prp22p with intron sequences at the 3' splice site is dependent on Prp16p and does not persist when release of mature mRNA from the spliceosome is blocked. Taken together, these results suggest that the 3' splice site of yeast introns is contacted much earlier than previously envisioned by a protein of approximately 120 kDa, and that a transient association of Prp22p with the 3' splice site occurs between the first and second catalytic steps. SN - 1362-4962 UR - https://www.unboundmedicine.com/medline/citation/10684925/Interaction_of_the_yeast_DExH_box_RNA_helicase_prp22p_with_the_3'_splice_site_during_the_second_step_of_nuclear_pre_mRNA_splicing_ L2 - https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/28.6.1313 DB - PRIME DP - Unbound Medicine ER -