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PMID: 6425802 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Mapping of psoralen cross-linked nucleotides in RNA.

Nucleic acids research ·Vol. 12 ·No. 7 ·1984-04-11 ·Pages 3405-23

Garrett-Wheeler E, Lockard RE, Kumar A

Abstract

A method is described for using the cross-linking reagent 4'-(hydroxy-methyl)-4,5',8-trimethylpsoralen (HMT) to map base paired regions and higher-order structure within RNA molecules. Applying this method to yeast tRNAPhe, we have specifically identified cross-links within the acceptor stem between U6 X U68, in the D-stem between C11 X C25, and in the T psi-stem between U50 X C63 and U52 X C63. We have also identified a unique cross-link between U8 X C48 which are trans pyrimidines in the core region due to tertiary interactions between U8:A14 and C48:G15. The precise point of cross-linking was deduced in every case by using purine-specific U2 ribonuclease along with cytidine-specific CL3 ribonuclease which will anomalously cleave after photoreversed pyrimidines. The ability to map the precise point of cross-linking should prove invaluable in identifying nucleotides in close proximity within the tertiary structure of other RNA molecules.

MeSH Terms

Base Composition Base Sequence Chemical Phenomena Chemistry Cross-Linking Reagents Furocoumarins Nucleic Acid Conformation Oligoribonucleotides Phosphorus Radioisotopes RNA, Transfer, Amino Acyl Ribonuclease T1 Trioxsalen/analogs & derivatives

Chemicals

Cross-Linking Reagents Furocoumarins Oligoribonucleotides Phosphorus Radioisotopes RNA, Transfer, Amino Acyl hydroxymethyltrioxsalen Ribonuclease T1 Trioxsalen

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Garrett-Wheeler E

Lockard R E

Kumar A

References (21)

21 references, click to expand

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/12.7.3405

Published

1984-04-11

Pages

3405-23

Language

English

Region

England

NLM ID

0411011

PMCID

PMC318754

Subset

Grants

NIGMS NIH HHS · GM 27692 · United States

NIGMS NIH HHS · GM 28985 · United States

NIGMS NIH HHS · GM 29488 · United States

External Links

PubMed source DOI: 10.1093/nar/12.7.3405 PMC Full Text View journal details

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Mapping of psoralen cross-linked nucleotides in RNA.

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