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PMID: 11226169 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Sm protein-Sm site RNA interactions within the inner ring of the spliceosomal snRNP core structure.

The EMBO journal ·Vol. 20 ·No. 1-2 ·2001-01-15 ·Pages 187-96

Urlaub H, Raker VA, Kostka S, Lührmann R

Abstract

Seven Sm proteins, E, F, G, D1, D2, D3 and B/B', assemble in a stepwise manner onto the single-stranded Sm site element (PuAU(4-6)GPu) of the U1, U2, U4 and U5 spliceosomal snRNAs, resulting in a doughnut-shaped core RNP structure. Here we show by UV cross-linking experiments using an Sm site RNA oligonucleotide (AAUUUUUGA) that several Sm proteins contact the Sm site RNA, with the most efficient cross-links observed for the G and B/B' proteins. Site-specific photo-cross-linking revealed that the G and B/B' proteins contact distinct uridines (in the first and third positions, respectively) in a highly position-specific manner. Amino acids involved in contacting the RNA are located at equivalent regions in both proteins, namely in loop L3 of the Sm1 motif, which has been predicted to jut into the hole of the Sm ring. Our results thus provide the first evidence that, within the core snRNP, multiple Sm protein-Sm site RNA contacts occur on the inner surface of the heptameric Sm protein ring.

MeSH Terms
Amino Acid Sequence Base Sequence Binding Sites Cross-Linking Reagents Models, Molecular Molecular Sequence Data Oligoribonucleotides/chemistry Protein Structure, Secondary RNA, Small Nuclear/chemistry Ribonucleoprotein, U1 Small Nuclear/chemistry Ribonucleoprotein, U2 Small Nuclear/chemistry Ribonucleoprotein, U4-U6 Small Nuclear/chemistry Ribonucleoprotein, U5 Small Nuclear/chemistry Ribonucleoproteins, Small Nuclear/chemistry Sequence Alignment Sequence Homology, Amino Acid Ultraviolet Rays
Chemicals
Cross-Linking Reagents Oligoribonucleotides RNA, Small Nuclear Ribonucleoprotein, U1 Small Nuclear Ribonucleoprotein, U2 Small Nuclear Ribonucleoprotein, U4-U6 Small Nuclear Ribonucleoprotein, U5 Small Nuclear Ribonucleoproteins, Small Nuclear
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Urlaub H
Max-Planck-Institute of Biophysical Chemistry, Department of Cellular Biochemistry, Am Fassberg 11, D-37077 Göttingen, Germany.
Raker V A
Kostka S
Lührmann R
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.1.187
Published
2001-01-15
Pages
187-96
Language
English
Region
England
NLM ID
8208664
PMCID
PMC140196
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.1.187 PMC Full Text View journal details
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