Premium
Complementation of DNA repair‐deficient Escherichia coli by the yeast Apn1 apurinic/apyrimidinic endonuclease gene
Author(s) -
Ramotar D.,
Popoff S. C.,
Demple B.
Publication year - 1991
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1991.tb01835.x
Subject(s) - biology , dna (apurinic or apyrimidinic site) lyase , escherichia coli , microbiology and biotechnology , dna repair , ap site , ap endonuclease , dna damage , dna , gene , genetics
Summary The Saccharomyces cerevisiae APN1 gene encoding an AP endonuclease/3′ ‐diesterase was engineered in vitro for expression in Escherichia coli. The expression vector directs the synthesis in E. coli of a M r 40500 protein that reacts with anti‐Apn1 antibodies and has the DNA‐repair activities characteristic of Apn1 isolated from yeast. A band corresponding to Apn1 was observed in DNA repair activity gels only with extracts of E. coli harbouring the APN1 expression plasmid. Expression of Apn1 conferred resistance to oxidants and alkylating agents in E. coli lacking exonuclease III and endonuclease IV. For H 2 O 2 damage, this rescue effect was correlated with the repair of oxidative lesions in the bacterial chromosome by the Apn1 protein. Thus, Apn1 can function in bacteria in a manner similar to its proposed multiple functions in yeast.