Document Detail


Heterogeneous nuclear RNA double-stranded regions probed in living HeLa cells by crosslinking with the psoralen derivative aminomethyltrioxsalen.
MedLine Citation:
PMID:  284397     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The psoralen derivative aminomethyltrioxsalen (AMT, 4'-aminomethyl-4,5',8-trimethylpsoralen) has been employed as a probe for heterogeneous nuclear RNA (hnRNA) double-stranded regions in experiments with living HeLa cells. hnRNA ribonucleoprotein (hnRNP) particles were purified from untreated or AMT-treated cells after irradiation with 365-nm light, and double-stranded hnRNA regions (dsRNA) were isolated by RNase A + T1 digestion of hnRNP, followed by preparative Cs2SO4 isopycnic centrifugation. The purified, hnRNP-derived dsRNA was then assayed for interstrand crosslinks by measurement of its "snapback" to RNase-resistant form after thermal denaturation. By this procedure, the amount of crosslinked dsRNA was found to be increased 3- to 7-fold in cells exposed to AMT in vivo. The levels of crosslinking in vivo compared favorably with those observed in model experiments with pure dsRNA in vitro. These results establish that double-stranded hnRNA regions exist in the living cell, and they further demonstrate that these base-paired regions are organized as rather accessible sites within the nucleus.
Authors:
J P Calvet; T Pederson
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Proceedings of the National Academy of Sciences of the United States of America     Volume:  76     ISSN:  0027-8424     ISO Abbreviation:  Proc. Natl. Acad. Sci. U.S.A.     Publication Date:  1979 Feb 
Date Detail:
Created Date:  1979-05-23     Completed Date:  1979-05-23     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7505876     Medline TA:  Proc Natl Acad Sci U S A     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  755-9     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Hela Cells
Hydrogen Bonding
Nucleic Acid Conformation
Nucleic Acid Precursors
Psoralens* / analogs & derivatives
RNA, Heterogeneous Nuclear*
RNA, Messenger
Trioxsalen* / analogs & derivatives,  pharmacology
Chemical
Reg. No./Substance:
0/Nucleic Acid Precursors; 0/Psoralens; 0/RNA, Heterogeneous Nuclear; 0/RNA, Messenger; 3902-71-4/Trioxsalen
Comments/Corrections

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