This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in ISI Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via ISI Web of Science (16) Citing Articles via Google Scholar Google Scholar Articles by Li, A. Articles by Tinland, B. Search for Related Content PubMed PubMed Citation Articles by Li, A. Articles by Tinland, B. Agricola Articles by Li, A. Articles by Tinland, B.

Repair of Damaged DNA by Arabidopsis Cell Extract

^a Institute of Plant Sciences, Eidgenössische Technishe Hochschule, Universitätsstr 2, CH-8092 Zürich, Switzerland
^b Friedrich Miescher Institute, P.O. Box 2543, CH-4002, Basel, Switzerland

⁴ To whom correspondence should be addressed. E-mail david.schuermann{at}fmi.ch; fax 41-61-697-39-76

All living organisms have to protect the integrity of their genomes from a wide range of genotoxic stresses to which they are inevitably exposed. However, understanding of DNA repair in plants lags far behind such knowledge in bacteria, yeast, and mammals, partially as a result of the absence of efficient in vitro systems. Here, we report the experimental setup for an Arabidopsis in vitro repair synthesis assay. The repair of plasmid DNA treated with three different DNA-damaging agents, UV light, cisplatin, and methylene blue, after incubation with whole-cell extract was monitored. To validate the reliability of our assay, we analyzed the repair proficiency of plants depleted in AtRAD1 activity. The reduced repair of UV light– and cisplatin-damaged DNA confirmed the deficiency of these plants in nucleotide excision repair. Decreased repair of methylene blue–induced oxidative lesions, which are believed to be processed by the base excision repair machinery in mammalian cells, may indicate a possible involvement of AtRAD1 in the repair of oxidative damage. Differences in sensitivity to DNA polymerase inhibitors (aphidicolin and dideoxy TTP) between plant and human cell extracts were observed with this assay.

This article has been cited by other articles:

				S. Takahashi, A. Sakamoto, S. Sato, T. Kato, S. Tabata, and A. Tanaka Roles of Arabidopsis AtREV1 and AtREV7 in Translesion Synthesis Plant Physiology, June 1, 2005; 138(2): 870 - 881. [Abstract] [Full Text] [PDF]

				J. Molinier, C. Ramos, O. Fritsch, and B. Hohn CENTRIN2 Modulates Homologous Recombination and Nucleotide Excision Repair in Arabidopsis PLANT CELL, June 1, 2004; 16(6): 1633 - 1643. [Abstract] [Full Text] [PDF]

				S. Kimura, Y. Tahira, T. Ishibashi, Y. Mori, T. Mori, J. Hashimoto, and K. Sakaguchi DNA repair in higher plants; photoreactivation is the major DNA repair pathway in non-proliferating cells while excision repair (nucleotide excision repair and base excision repair) is active in proliferating cells Nucleic Acids Res., May 18, 2004; 32(9): 2760 - 2767. [Abstract] [Full Text] [PDF]

				F. Lincker, G. Philipps, and M.-E. Chaboute UV-C response of the ribonucleotide reductase large subunit involves both E2F-mediated gene transcriptional regulation and protein subcellular relocalization in tobacco cells Nucleic Acids Res., February 27, 2004; 32(4): 1430 - 1438. [Abstract] [Full Text] [PDF]

				Z. Liu, S.-W. Hong, M. Escobar, E. Vierling, D. L. Mitchell, D. W. Mount, and J. D. Hall Arabidopsis UVH6, a Homolog of Human XPD and Yeast RAD3 DNA Repair Genes, Functions in DNA Repair and Is Essential for Plant Growth Plant Physiology, July 1, 2003; 132(3): 1405 - 1414. [Abstract] [Full Text] [PDF]