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A more recent version of this article appeared on June 13, 2003
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M302585200v1
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Papers In Press, published online ahead of print April 11, 2003
J. Biol. Chem, 10.1074/jbc.M302585200
Submitted on March 13, 2003
Revised on April 9, 2003
Accepted on April 11, 2003

DNA loop repair by Escherichia coli cell extracts

Woei-horng Fang, Bo-Jeng Wang, Chiang-Hui Wang, Su-Jan Lee, Yu-Ting Chang, Yi-Kuang Chuang, and Chun-Nan Lee

School of Medical Technology, College of Medicine, National Taiwan University, Taipei, Taiwan 100-02

Corresponding Author: whfang{at}ha.mc.ntu.edu.tw

The nick-directed DNA repair efficiency of a set of M13mp18 derived heteroduplexes containing 8-, 12-, 16-, 22-, 27-, 45-, and 429-nucleotide loops was determined by in vitro assay. Unpaired nucleotides of each heteroduplexes reside within overlapping recognition sites for two restriction endonucleases, permitting independent evaluation of repair occurring on either DNA strand. Our results show that a strand break located either on 3’ or 5’ to the loop is suffice to direct heterology repair to the nicked strand in Escherichia coli extracts. Strand-specific repair by this system requires Mg2+ and the four dNTPs and is equally efficient on insertions and deletions. This activity is distinct from MutHLS mismatch repair pathway. Strand specificity and repair efficiency are largely independent of the GATC methylation state of the DNA and presence of the products of mismatch repair genes mutH, mutL, and mutS. This study provides evidence for a loop repair pathway in E. coli that is distinct from conventional mismatch repair.


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