Repair of psoralen crosslinks in Escherichia coli: In vitro studies with the RecA protein and (A)BC excinuclease
RecA,a key protein for recombination in Escherichia coli (E. Coli), was utilized to insert short psoralen-monoadducted oligonucleotides into a duplex plasmid molecule through a variation of the homologous pairing reaction. Covalent psoralen crosslinks could then be formed between the oligonucleotide and its complement within the plasmid. The efficiency of the overall process was found to depend upon the nucleoside triphosphate cofactor, the extent of homology, oligonucleotide length, and psoralen location. Deproteinized complexes were also observed to retain a certain degree of unwinding of the plasmid due to the crosslinked oligonucleotides. These three-stranded complexes were used as model substrates for the study of the action mechanism of excinuclease, the enzyme responsible for nucleotide excision repair in E. coli. 290 refs., 24 figs.
- Research Organization:
- Lawrence Berkeley Lab., CA (USA)
- Sponsoring Organization:
- DOE/ER
- DOE Contract Number:
- AC03-76SF00098
- OSTI ID:
- 7192240
- Report Number(s):
- LBL-28930; ON: DE90011337
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550700* -- Microbiology
59 BASIC BIOLOGICAL SCIENCES
ANTICOAGULANTS
BACTERIA
BIOLOGICAL FUNCTIONS
BIOLOGICAL PATHWAYS
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CELL CONSTITUENTS
COUMARINS
DNA
DNA POLYMERASES
DNA REPAIR
DRUGS
EFFICIENCY
ENZYMES
ESCHERICHIA COLI
EXCISION REPAIR
FUNCTIONS
GENE RECOMBINATION
HEMATOLOGIC AGENTS
HETEROCYCLIC COMPOUNDS
IN VITRO
LIGASES
MICROORGANISMS
NUCLEIC ACIDS
NUCLEOSIDES
NUCLEOTIDES
NUCLEOTIDYLTRANSFERASES
OLIGONUCLEOTIDES
ORGANIC COMPOUNDS
ORGANIC OXYGEN COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PLASMIDS
POLYMERASES
PROTEINS
PSORALEN
RECOVERY
REPAIR
RIBOSIDES
TRANSFERASES