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PMID: 2529425 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

U1 small nuclear ribonucleoprotein particle-specific proteins interact with the first and second stem-loops of U1 RNA, with the A protein binding directly to the RNA independently of the 70K and Sm proteins.

Molecular and cellular biology ·Vol. 9 ·No. 8 ·1989-08-00 ·Pages 3360-8

Patton JR, Habets W, van Venrooij WJ, Pederson T

Abstract

The U1 small nuclear ribonucleoprotein particle (U1 snRNP), a cofactor in pre-mRNA splicing, contains three proteins, termed 70K, A, and C, that are not present in the other spliceosome-associated snRNPs. We studied the binding of the A and C proteins to U1 RNA, using a U1 snRNP reconstitution system and an antibody-induced nuclease protection technique. Antibodies that reacted with the A and C proteins induced nuclease protection of the first two stem-loops of U1 RNA in reconstituted U1 snRNP. Detailed analysis of the antibody-induced nuclease protection patterns indicated the existence of relatively long-range protein-protein interactions in the U1 snRNP, with the 5' end of U1 RNA and its associated specific proteins interacting with proteins bound to the Sm domain near the 3' end. UV cross-linking experiments in conjunction with an A-protein-specific antibody demonstrated that the A protein bound directly to the U1 RNA rather than assembling in the U1 snRNP exclusively via protein-protein interactions. This conclusion was supported by additional experiments revealing that the A protein could bind to U1 RNA in the absence of bound 70K and Sm core proteins.

MeSH Terms
Autoantibodies Base Sequence Humans Micrococcal Nuclease Molecular Sequence Data Nucleic Acid Conformation Oligoribonucleotides/analysis Protein Binding RNA, Small Nuclear/metabolism,radiation effects Ribonucleoproteins/metabolism,radiation effects Ribonucleoproteins, Small Nuclear Sepharose/analogs & derivatives Ultraviolet Rays
Chemicals
Autoantibodies Oligoribonucleotides RNA, Small Nuclear Ribonucleoproteins Ribonucleoproteins, Small Nuclear Staphylococcal protein A-sepharose Sepharose Micrococcal Nuclease
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Patton J R
Cell Biology Group, Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts 01545.
Habets W
van Venrooij W J
Pederson T
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.9.8.3360-3368.1989
Published
1989-08-00
Pages
3360-8
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362381
Subset
IM
Grants
NIGMS NIH HHS · GM-11399 · United States
NIGMS NIH HHS · GM-21595-14 · United States
External Links
PubMed source DOI: 10.1128/mcb.9.8.3360-3368.1989 PMC Full Text View journal details
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