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PMID: 8076607 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The association of the U1-specific 70K and C proteins with U1 snRNPs is mediated in part by common U snRNP proteins.

The EMBO journal ·Vol. 13 ·No. 17 ·1994-09-01 ·Pages 4113-25

Nelissen RL, Will CL, van Venrooij WJ, Lührmann R

Abstract

The U1 small nuclear ribonucleoprotein particle (snRNP)-specific 70K and A proteins are known to bind directly to stem-loops of the U1 snRNA, whereas the U1-C protein does not bind to naked U1 snRNA, but depends on other U1 snRNP protein components for its association. Focusing on the U1-70K and U1-C proteins, protein-protein interactions contributing to the association of these particle-specific proteins with the U1 snRNP were studied. Immunoprecipitation of complexes formed after incubation of naked U1 snRNA or purified U1 snRNPs lacking their specific proteins (core U1 snRNP) with in vitro translated U1-C protein, revealed that both common snRNP proteins and the U1-70K protein are required for the association of U1-C with the U1 snRNP. Binding studies with various in vitro translated U1-70K mutants demonstrated that the U1-70K N-terminal domain is necessary and sufficient for the interaction of U1-C with core U1 snRNPs. Surprisingly, several N-terminal fragments of the U1-70K protein, which lacked the U1-70K RNP-80 motif and did not bind naked U1 RNA, associated stably with core U1 snRNPs. This suggests that a new U1-70K binding site is generated upon association of common U1 snRNP proteins with U1 RNA. The interaction between the N-terminal domain of U1-70K and the core RNP domain was specific for the U1 snRNP; stable binding was not observed with core U2 or U5 snRNPs, suggesting essential structural differences among snRNP core domains. Evidence for direct protein-protein interactions between U1-specific proteins and common snRNP proteins was supported by chemical crosslinking experiments using purified U1 snRNPs. Individual crosslinks between the U1-70K and the common D2 or B'/B protein, as well as between U1-C and B'/B, were detected. A model for the assembly of U1 snRNP is presented in which the complex of common proteins on the RNA backbone functions as a platform for the association of the U1-specific proteins.

MeSH Terms
Autoantigens/metabolism Binding Sites Cross-Linking Reagents DNA Mutational Analysis Humans Models, Biological Precipitin Tests Protein Binding RNA, Small Nuclear/metabolism RNA-Binding Proteins/metabolism Ribonucleoprotein, U1 Small Nuclear/metabolism,ultrastructure Ribonucleoproteins, Small Nuclear Sequence Deletion Structure-Activity Relationship snRNP Core Proteins
Chemicals
Autoantigens Cross-Linking Reagents RNA, Small Nuclear RNA-Binding Proteins Ribonucleoprotein, U1 Small Nuclear Ribonucleoproteins, Small Nuclear snRNP Core Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Nelissen R L
Department of Biochemistry, University of Nijmegen, The Netherlands.
Will C L
van Venrooij W J
Lührmann R
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37 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1994-09-01
Pages
4113-25
Language
English
Region
England
NLM ID
8208664
PMCID
PMC395334
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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