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PMID: 2974535 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

An in vitro interaction between the human U3 snRNP and 28S rRNA sequences near the alpha-sarcin site.

Nucleic acids research ·Vol. 16 ·No. 22 ·1988-11-25 ·Pages 10493-509

Parker KA, Bruzik JP, Steitz JA

Abstract

Model transcripts containing mammalian pre-rRNA sequences were incubated with a HeLa cell extract, digested with T1 RNase, and immunoprecipitated with anti-(U3)RNP or control antibodies. Two overlapping fragments derived from the 3' domain of human 28S rRNA were specifically immunoprecipitated although transcripts which spanned the transcription initiation site, the ETS processing site, the 5' end of 18S, and both termini of 5.8S yielded no protected fragments. The sequence of these fragments was determined using a novel technique in which the [32P]-labeled fragment was co-finger-printed with [3H]-labeled total transcript serving as an internal marker. The fragments immunoprecipitated derive from nucleotides 4570-4590 and 4575-4590 of human 28S and are adjacent to the alpha-sarcin site. Protection most likely involves the U3 RNA since it is sensitive to pretreatment of the extract with micrococcal nuclease. Complementarity between U3 and this rRNA region is phylogenetically conserved in species ranging from human to S. cerevisiae. The possible significance of this finding is discussed.

MeSH Terms
Antibodies Base Composition Base Sequence Endoribonucleases Fungal Proteins/genetics HeLa Cells/metabolism Humans Molecular Sequence Data Nucleic Acid Conformation Plasmids Protein Synthesis Inhibitors RNA Precursors/genetics RNA, Ribosomal/genetics RNA, Ribosomal, 28S/genetics,metabolism Ribonuclease T1 Ribonucleoproteins/genetics,metabolism Ribonucleoproteins, Small Nuclear Transcription, Genetic
Chemicals
Antibodies Fungal Proteins Protein Synthesis Inhibitors RNA Precursors RNA, Ribosomal RNA, Ribosomal, 28S Ribonucleoproteins Ribonucleoproteins, Small Nuclear alpha-sarcin Endoribonucleases Ribonuclease T1
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Parker K A
Department of Molecular Biophysics and Biochemistry, Howard Hughes Medical Institute, Yale University Medical School, New Haven, CT 06510.
Bruzik J P
Steitz J A
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/16.22.10493
Published
1988-11-25
Pages
10493-509
Language
English
Region
England
NLM ID
0411011
PMCID
PMC338920
Subset
IM
Grants
NIGMS NIH HHS · GM 26154 · United States
External Links
PubMed source DOI: 10.1093/nar/16.22.10493 PMC Full Text View journal details
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