Complementation of DNA repair‐deficient Escherichia coli by the yeast Apn1 apurinic/apyrimidinic endonuclease gene

D. Ramotar; S. C. Popoff; B. Demple

doi:10.1111/j.1365-2958.1991.tb01835.x

Complementation of DNA repair‐deficient Escherichia coli by the yeast Apn1 apurinic/apyrimidinic endonuclease gene

D. Ramotar, S. C. Popoff, B. Demple^*

^*Corresponding author for this work

Harvard University

Research output: Contribution to journal › Article › peer-review

45 Citations (Scopus)

Abstract

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 H₂O₂ 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.

Original language	English
Pages (from-to)	149-155
Number of pages	7
Journal	Molecular Microbiology
Volume	5
Issue number	1
DOIs	https://doi.org/10.1111/j.1365-2958.1991.tb01835.x
Publication status	Published - Jan 1991
Externally published	Yes

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title = "Complementation of DNA repair‐deficient Escherichia coli by the yeast Apn1 apurinic/apyrimidinic endonuclease gene",

abstract = "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 Mr 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 H2O2 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.",

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AB - 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 Mr 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 H2O2 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.

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Complementation of DNA repair‐deficient Escherichia coli by the yeast Apn1 apurinic/apyrimidinic endonuclease gene

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