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Repair of psoralen crosslinks in Escherichia coli: In vitro studies with the RecA protein and (A)BC excinuclease

Technical Report ·
DOI:https://doi.org/10.2172/7192240· OSTI ID:7192240

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