Analysis of ultraviolet-induced RNA-RNA cross-links: A means for probing RNA structure-function relationships

Andrea D. Branch, Bonnie J. Benenfeld, Cynthia P. Paul, Hugh D. Robertson

Research output: Contribution to journalArticlepeer-review

36 Scopus citations

Abstract

This chapter presents analysis of ultraviolet-induced RNA–RNA cross-links. To obtain maximum information about local bonding patterns, the UV-induced cross-link must be mapped to the nucleotide level. Direct RNA analysis is needed. However, if cross-links are to be used only to determine which portions of the RNA are close together in the three-dimensional structure, indirect techniques, such as primer extension, may be employed. The mapping of an extensive series of RNA–RNA cross-links introduced into rRNA has led to a detailed three-dimensional picture of 16 S rRNA. While additional procedures are usually required to locate a UV-induced bond in a large RNA molecule, a cross-link in an RNA smaller than 1000 nt can often be detected directly by a change the cross-link makes in the characteristic pattern of ribonuclease-resistant oligonucleotides. Primer extension is useful for mapping cross-links, especially when employed in combination with direct RNA analysis.

Original languageEnglish
Pages (from-to)418-442
Number of pages25
JournalMethods in Enzymology
Volume180
Issue numberC
DOIs
StatePublished - 1 Jan 1989

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