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.
- Research Organization:
- Univ. of California, Berkeley, CA (United States)
- DOE Contract Number:
- W-7405-ENG-48
- OSTI ID:
- 6267359
- Journal Information:
- Cell; (United States), Vol. 32
- Country of Publication:
- United States
- Language:
- English
Similar Records
Reaction of psoralen with RNA: specificity and use as a probe for secondary-structure analysis
Specificity of the photoreaction of 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen with ribonucleic acid. Identificaton of reactive sites in Escherichia coli phenylalanine-accepting transfer ribonucleic acid
Related Subjects
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)