Reaction of psoralen with RNA: specificity and use as a probe for secondary-structure analysis
A variety of techniques has been used to study how psoralen and its derivatives react with RNA. This information has then been used to analyze the secondary structure of different ribosomal RNAs. Paper electrophoresis at pH 3.5 and 8.8 and HPLC has been used to get high-resolution separation of RNA-psoralen adducts. The separated adducts have been analyzed and shown to be primarily uridine adducts with the psoralen reacted at the furan end. The stereochemistry of the major adducts was determined by NMR. The effect of structural transitions on the number and type of adducts was found for several polymers. The effect of psoralen structure on cross linking ability was analyzed. Charged derivatives formed monoadducts very efficiently but did not produce the level of crosslinking obtainable with lower levels of reaction with uncharged derivatives. Secondary structure analysis of D. melanogaster 5S RNA yielded two definite and two tentative crosslinks which support the generally accepted models for 5S structure. Analysis of E. coli 16S RNA by gel techniques yielded 13 cross-links. Evidence is also presented for an interaction between eukaryotic mRNA (5' cap structure) and 18S RNA (hypermodified base am psi) which serves a function analogous to the Shine-Dalgarno sequence in pro karyotes.
- Research Organization:
- Lawrence Berkeley Lab., CA (USA)
- DOE Contract Number:
- AC03-76SF00098
- OSTI ID:
- 6871694
- Report Number(s):
- LBL-14991; ON: DE83003588
- Resource Relation:
- Other Information: Portions of document are illegible. Thesis
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PSORALEN
PHOTOCHEMICAL REACTIONS
RNA
MOLECULAR STRUCTURE
ADDUCTS
CHROMATOGRAPHY
CROSS-LINKING
DROSOPHILA
ELECTROPHORESIS
ESCHERICHIA COLI
EXPERIMENTAL DATA
RIBOSOMES
ANIMALS
ANTICOAGULANTS
ARTHROPODS
BACTERIA
CELL CONSTITUENTS
CHEMICAL REACTIONS
COUMARINS
DATA
DIPTERA
DRUGS
FLIES
FRUIT FLIES
HEMATOLOGIC AGENTS
HETEROCYCLIC COMPOUNDS
INFORMATION
INSECTS
INVERTEBRATES
MICROORGANISMS
NUCLEIC ACIDS
NUMERICAL DATA
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
ORGANOIDS
POLYMERIZATION
SEPARATION PROCESSES
550200* - Biochemistry