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Title: Structure of E. coli 16S RNA elucidated by psoralen crosslinking

E. coli 16S RNA in solution was photoreacted with hydroxymethyltrimethylpsoralen and long wave ultraviolet light. Positions of crosslinks were determined to high resolution by partially digesting the RNA with T/sub 1/ RNase, separating the crosslinked fragments by two-dimensional gel electrophoresis, reversing the crosslink, and sequencing the separated fragments. This method yielded the locations of crosslinks to +/-15 nucleotides. Even finer placement has been made on the basis of our knowledge of psoralen reactivity. Thirteen unique crosslinks were mapped. Seven crosslinks confirmed regions of secondary structure which had been predicted in published phylogenetic models, three crosslinks discriminated between phylogenetic models, and three proved the existence of new structures. The new structures were all long-range interactions which appear to be in dynamic equilibrium with local secondary structure. Because this technique yields direct information about the secondary structure of large RNAs, it should prove invaluable in studying the structure of other RNAs of all sizes.
Authors:
;
Publication Date:
OSTI Identifier:
6267359
DOE Contract Number:
W-7405-ENG-48
Resource Type:
Journal Article
Resource Relation:
Journal Name: Cell; (United States); Journal Volume: 32
Research Org:
Univ. of California, Berkeley
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; NUCLEOTIDES; CROSS-LINKING; PSORALEN; PHOTOCHEMICAL REACTIONS; RNA; BIOLOGICAL RADIATION EFFECTS; STRUCTURAL CHEMICAL ANALYSIS; ELECTROPHORESIS; ESCHERICHIA COLI; GENETIC MAPPING; MATHEMATICAL MODELS; MOLECULAR STRUCTURE; ULTRAVIOLET RADIATION; ANTICOAGULANTS; BACTERIA; BIOLOGICAL EFFECTS; CHEMICAL REACTIONS; COUMARINS; DRUGS; ELECTROMAGNETIC RADIATION; HEMATOLOGIC AGENTS; HETEROCYCLIC COMPOUNDS; MAPPING; MICROORGANISMS; NUCLEIC ACIDS; ORGANIC COMPOUNDS; ORGANIC OXYGEN COMPOUNDS; POLYMERIZATION; RADIATION EFFECTS; RADIATIONS 550200* -- Biochemistry; 560111 -- Radiation Effects on Biochemicals-- In Vitro-- (-1987)