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Molecular and Cellular Biology, December 2000, p. 8836-8844, Vol. 20, No. 23
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

DNA Integration by Ty Integrase in yku70 Mutant Saccharomyces cerevisiae Cells

Markus Kiechle,1,2 Anna A. Friedl,1,3 Palaniyandi Manivasakam,2 Friederike Eckardt-Schupp,1 and Robert H. Schiestl2,*

Institute of Radiobiology, GSF Research Center, D-85764 Neuherberg,1 and Radiobiological Institute, University of Munich, D-80336 Munich,3 Germany, and Department of Cancer Cell Biology, Harvard School of Public Health, Boston, Massachusetts 021152

Received 21 June 2000/Returned for modification 25 August 2000/Accepted 5 September 2000

In the present work we examined nonhomologous integration of plasmid DNA in a yku70 mutant. Ten of 14 plasmids integrated as composite elements, including Ty sequences probably originating from erroneous strand-switching and/or priming events. Three additional plasmids integrated via Ty integrase without cointegrating Ty sequences, as inferred from 5-bp target site duplication and integration site preferences. Ty integrase-mediated integration of non-Ty DNA has never been observed in wild-type cells, although purified integrase is capable of using non-Ty DNA as a substrate in vitro. Hence our data implicate yKu70 as the cellular function preventing integrase from accepting non-Ty DNA as a substrate.

* Corresponding author. Mailing address: Department of Cancer Cell Biology, Harvard School of Public Health, 665 Huntington Ave., Boston, MA 02115. Phone: (617) 432-4410. Fax: (617) 432-2059. E-mail: schiestl{at}hsph.harvard.edu.

Molecular and Cellular Biology, December 2000, p. 8836-8844, Vol. 20, No. 23
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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