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PMID: 2771939 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

U3 small nuclear RNA can be psoralen-cross-linked in vivo to the 5' external transcribed spacer of pre-ribosomal-RNA.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 86 ·No. 17 ·1989-09-00 ·Pages 6523-7

Maser RL, Calvet JP

Abstract

U3 small nuclear RNA is hydrogen-bonded to high molecular weight nucleolar RNA and can be isolated from greater than 60S pre-ribosomal ribonucleoprotein particles, suggesting that it is involved in processing of ribosomal RNA precursors (pre-rRNA) or in ribosome biogenesis. Here we have used in vivo psoralen cross-linking to identify the region of pre-rRNA interacting with U3 RNA. Quantitative hybridization selection/depletion experiments with clones of rRNA-encoding DNA (rDNA) and cross-linked nuclear RNA showed that all of the cross-linked U3 RNA was associated with a region that includes the external transcribed spacer (ETS) at the 5' end of the human rRNA precursor. To further identify the site of interaction within the approximately 3.7-kilobase ETS, Southern blots of rDNA clones were sandwich-hybridized with cross-linked RNA and then probed for cross-linked U3 RNA. These experiments showed that U3 RNA was cross-linked to a 258-base sequence between nucleotides +438 and +695, just downstream of the ETS early cleavage site (+414). The localization of U3 to this region of the rRNA precursor was not expected from previous models for a base-paired U3-rRNA interaction and suggests that U3 plays a role in the initial pre-rRNA processing event.

MeSH Terms
Blotting, Southern Cloning, Molecular Cross-Linking Reagents DNA, Ribosomal/genetics,metabolism Furocoumarins/pharmacology HeLa Cells/metabolism Humans Multigene Family Nucleic Acid Hybridization RNA Precursors/genetics,metabolism RNA, Small Nuclear/metabolism Transcription, Genetic
Chemicals
Cross-Linking Reagents DNA, Ribosomal Furocoumarins RNA Precursors RNA, Small Nuclear
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Maser R L
Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City 66103.
Calvet J P
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.86.17.6523
Published
1989-09-00
Pages
6523-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC297876
Subset
IM
Grants
NIGMS NIH HHS · GM36547 · United States
External Links
PubMed source DOI: 10.1073/pnas.86.17.6523 PMC Full Text View journal details
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