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PMID: 8608445 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The canonical GU dinucleotide at the 5' splice site is recognized by p220 of the U5 snRNP within the spliceosome.

RNA (New York, N.Y.) ·Vol. 2 ·No. 3 ·1996-03-00 ·Pages 213-25

Reyes JL, Kois P, Konforti BB, Konarska MM

Abstract

Specific recognition of the 5' splice site (5'SS) by the spliceosome components was studied using a simple in vitro system in which a short 5'SS RNA oligonucleotide specifically induces the assembly of snRNP particles into spliceosome-like complexes and actively participates in a trans-splicing reaction. Short-range cross-liking demonstrates that a U5 snRNP protein component, p220 (the human analogue of the yeast Prp8) specifically interacts with the invariant GU dinucleotide at the 5' end of the intron. The GU:p220 interaction can be detected in the functional splicing complex B. Although p220 has been known to contact several nucleotides around the 5' splice junction, the p220:GU dinucleotide interaction described here is remarkably specific. Consistent with the high conservation of the GU, even minor modifications of this element affect recognition of the 5'SS RNA by p220. Substitution of uridine at the GU with base analogues containing a large methyl or iodo group, but not a smaller flouro group at base position 5, interferes with association of 5'SS RNA with snRNP complexes and their functional participation in splicing.

MeSH Terms
Base Sequence Conserved Sequence Cross-Linking Reagents Humans Introns Molecular Sequence Data Oligoribonucleotides/metabolism Protein Binding RNA Precursors/metabolism RNA Splicing/physiology Ribonucleoprotein, U5 Small Nuclear/metabolism Ribonucleoproteins, Small Nuclear/metabolism Spliceosomes/metabolism Thiouridine/metabolism Ultraviolet Rays
Chemicals
Cross-Linking Reagents Oligoribonucleotides RNA Precursors Ribonucleoprotein, U5 Small Nuclear Ribonucleoproteins, Small Nuclear Thiouridine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Reyes J L
Rockefeller University, New York, New York 10021, USA.
Kois P
Konforti B B
Konarska M M
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
Published
1996-03-00
Pages
213-25
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1369364
Subset
IM
Grants
NIGMS NIH HHS · GM49044 · United States
External Links
PubMed source PMC Full Text View journal details
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