Linked Life Data

1531654

Source:http://linkedlifedata.com/resource/pubmed/id/1531654

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
7
pubmed:dateCreated
1992-4-2
pubmed:abstractText
Secondary structures for all five spliceosomal small nuclear (sn) RNAs (U1, U2, U4, U5, and U6 snRNAs) have been derived from phylogenetic, biochemical, and genetic data, but tertiary structure information has been more difficult to obtain. Here we have used the general cross-linking reagent nitrogen mustard (bis-(2-chloroethyl)methylamine) to explore the tertiary conformation of naked U1 snRNA. We detected two intramolecularly cross-linked U1 species (X-U1#1 and X-U1#2) after cross-linking of deproteinized HeLa nuclear extract. We determined the cross-linked sites and found that X-U1#1 is cross-linked between the C82-A85 and U129, while X-U1#2 is cross-linked between U105-G108 and A118. We then tested the ability of these two cross-linked species to bind small nuclear ribonucleo-protein (snRNP) proteins in vitro (in HeLa nuclear extract or S100) and in vivo (in Xenopus oocytes). Both X-U1#1 and X-U1#2 were found to reconstitute efficiently in vitro and in vivo, as judged by immunoprecipitation with antibodies specific for Sm and U1-specific proteins. Our data suggest that (i) the Sm-binding site lies on the surface of the native U1 snRNP, since the cross-link in X-U1#1 involves the Sm-binding site but does not block snRNP assembly, and (ii) U1 snRNA may adopt the correct tertiary conformation even in the absence of U1 snRNP proteins.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
5
pubmed:volume
267
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
4503-7
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1992
pubmed:articleTitle
Cross-linking of U1 snRNA using nitrogen mustard. Evidence for higher order structure.
pubmed:affiliation
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06510.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.