OPEN ACCESS ARTICLE

This Article Free via Open Access: OA OA Full Text Full Text (PDF) OA All Versions of this Article: 146/4/1941    most recent pp.107.110189v1 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 (2) Citing Articles via Google Scholar Google Scholar Articles by Teranishi, M. Articles by Hidema, J. Search for Related Content PubMed PubMed Citation Articles by Teranishi, M. Articles by Hidema, J. Agricola Articles by Teranishi, M. Articles by Hidema, J.

ENVIRONMENTAL STRESS AND ADAPTATION TO STRESS

The Native Cyclobutane Pyrimidine Dimer Photolyase of Rice Is Phosphorylated^1,[C],[OA]

Department of Environmental Life Sciences (M.T., K.N., J.H.) and Department of Biomolecular Sciences (K.Y.), Graduate School of Life Sciences, Tohoku University, Sendai 980–8577, Japan; and Department of Analytical and Biophysical Chemistry, Faculty of Medical and Pharmaceutical Sciences, Kumamoto University, Kumamoto 862–0973, Japan (H.M.)

The cyclobutane pyrimidine dimer (CPD) is a major type of DNA damage induced by ultraviolet B (UVB) radiation. CPD photolyase, which absorbs blue/UVA light as an energy source to monomerize dimers, is a crucial factor for determining the sensitivity of rice (Oryza sativa) to UVB radiation. Here, we purified native class II CPD photolyase from rice leaves. As the final purification step, CPD photolyase was bound to CPD-containing DNA conjugated to magnetic beads and then released by blue-light irradiation. The final purified fraction contained 54- and 56-kD proteins, whereas rice CPD photolyase expressed from Escherichia coli was a single 55-kD protein. Western-blot analysis using anti-rice CPD photolyase antiserum suggested that both the 54- and 56-kD proteins were the CPD photolyase. Treatment with protein phosphatase revealed that the 56-kD native rice CPD photolyase was phosphorylated, whereas the E. coli-expressed rice CPD photolyase was not. The purified native rice CPD photolyase also had significantly higher CPD photorepair activity than the E. coli-expressed CPD photolyase. According to the absorption, emission, and excitation spectra, the purified native rice CPD photolyase possesses both a pterin-like chromophore and an FAD chromophore. The binding activity of the native rice CPD photolyase to thymine dimers was higher than that of the E. coli-expressed CPD photolyase. These results suggest that the structure of the native rice CPD photolyase differs significantly from that of the E. coli-expressed rice CPD photolyase, and the structural modification of the native CPD photolyase leads to higher activity in rice.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Mika Teranishi (tera{at}ige.tohoku.ac.jp).

^[C] Some figures in this article are displayed in color online but in black and white in the print edition.

^[OA] Open Access articles can be viewed online without a subscription.

www.plantphysiol.org/cgi/doi/10.1104/pp.107.110189

^* Corresponding author; e-mail tera{at}ige.tohoku.ac.jp.

Received October 4, 2007; accepted January 19, 2008; published January 30, 2008.

This article has been cited by other articles:

				K. Hitomi, L. DiTacchio, A. S. Arvai, J. Yamamoto, S.-T. Kim, T. Todo, J. A. Tainer, S. Iwai, S. Panda, and E. D. Getzoff Functional motifs in the (6-4) photolyase crystal structure make a comparative framework for DNA repair photolyases and clock cryptochromes PNAS, April 28, 2009; 106(17): 6962 - 6967. [Abstract] [Full Text] [PDF]