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Applied and Environmental Microbiology, October 2003, p. 6216-6224, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.6216-6224.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Nuclease Activity of Saccharomyces cerevisiae Mre11 Functions in Targeted Nucleotide Alteration

Li Liu, Michael Usher,{dagger} Yiling Hu, and Eric B. Kmiec*

Department of Biological Sciences, Delaware Biotechnology Institute, University of Delaware, Newark, Delaware 19711

Received 29 May 2003/ Accepted 7 July 2003

Oligonucleotides can be used to direct site-specific changes in genomic DNA through a process in which mismatched base pairs in the oligonucleotide and the target DNA are created. The mechanism by which these complexes are developed and resolved is being studied by using Saccharomyces cerevisiae as a model system. Genetic analyses have revealed that in all likelihood the reaction occurs in two phases: DNA pairing and DNA repair. While the former phase involves strand assimilation, the latter phase likely involves an endonucleolytic processing step that leads to joint resolution. In this study, we established the importance of a functioning MRE11 gene in the overall reaction, as yeast strains deficient in MRE11 exhibited severely reduced activity. The activity could be rescued by complementation with wild-type MRE11 genes but not with MRE11 alleles lacking the nuclease function. Taken together, the data suggest that Mre11 provides nuclease activity for targeted nucleotide exchange, a process that could be used to reengineer yeast genes.

* Corresponding author. Mailing address: Department of Biological Sciences, University of Delaware, Delaware Biotechnology Institute, 15 Innovation Way, Newark, DE 19711. Phone: (302) 831-3420. Fax: (302) 831-3427. E-mail: ekmiec{at}udel.edu.

{dagger} Present address: School of Medicine, University of Michigan, Ann Arbor, MI 48104.

Applied and Environmental Microbiology, October 2003, p. 6216-6224, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.6216-6224.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Wu, X.-S., Xin, L., Yin, W.-X., Shang, X.-Y., Lu, L., Watt, R. M., Cheah, K. S. E., Huang, J.-D., Liu, D.-P., Liang, C.-C. (2005). Increased efficiency of oligonucleotide-mediated gene repair through slowing replication fork progression. Proc. Natl. Acad. Sci. USA 102: 2508-2513 [Abstract] [Full Text]  


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