Similarity among the Drosophila (6-4)photolyase, a human photolyase homolog, and the DNA photolyase-blue-light photoreceptor family

Science

Volumn 272, Issue 5258, 1996, Pages 109-112

  Todo, Takeshi ^b   Ryo, Haruko ^b   Yamamoto, Kazuo ^b   Toh, Hiroyuki ^b   Inui, Taiichiro ^b   Ayaki, Hitoshi ^b   Nomura, Taisei ^b   Ikenaga, Mituo ^b  

^a KYOTO UNIVERSITY   (Japan)

^b NONE

Author keywords

[No Author keywords available]

Indexed keywords

CYCLOBUTANE DERIVATIVE; DEOXYRIBODIPYRIMIDINE PHOTOLYASE; PYRIMIDINE DIMER; PYRIMIDINONE DERIVATIVE;

AMINO ACID SEQUENCE; ARTICLE; CONTROLLED STUDY; DNA DAMAGE; DROSOPHILA MELANOGASTER; ENZYME ANALYSIS; MOLECULAR CLONING; NONHUMAN; PHOTORECEPTOR; PRIORITY JOURNAL; ULTRAVIOLET RADIATION;

EID: 0029914630     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.272.5258.109     Document Type: Article

References (29)

1. W. Harm, Biological Effects of Ultraviolet Radiation (Cambridge Univ. Press, Cambridge, 1980); E C. Freidberg, G. C. Walker, W. Siede, DNA Repair and Mutagenesis (ASM Press, Washington, DC, 1995).
2. W. Harm, Biological Effects of Ultraviolet Radiation (Cambridge Univ. Press, Cambridge, 1980); E C. Freidberg, G. C. Walker, W. Siede, DNA Repair and Mutagenesis (ASM Press, Washington, DC, 1995).
3. D. E. Brash, Photochem. Photobiol. 48, 59 (1988)
4. D. L. Mitchell and R. S. Nairn, ibid. 49, 805 (1989).
5. G B. Sancar, Mutat. Res 236, 147 (1990).
6. A. Sancar, Biochemistry 33, 2 (1994)
7. T Todo and H. Ryo, Mutat. Res 273, 85 (1992).
8. T. Todo et al., Nature 361, 371 (1993).
9. 2 of UV light from a germicidal lamp and then illuminated for 15 min with visible light at a distance of 24 cm from a FL15D lamp (National) with a 7-mm thick soft glass filter (PR treatment). Surviving cells were collected from the plates the next day and were subjected to a second round of selection with UV plus PR treatment. After 13 rounds of selection, four single colonies were selected from the illuminated plate, and their UV sensitivity with or without PR treatment was determined Plasmid DNA was isolated from one of the light-dependent UV-resistant colonies and designated pDm64PR.
10. T. Todo et al., data not shown.
11. T. Todo et al. Mutat Res. 315 213 (1994).
12. 2)] was labeled with 500 mg of photobiotin (Vector Laboratory) by irradiation in an ice bath 10 cm below a sunlamp (Toshiba, FL20SE) with a UV-34 filter (Hoya, Tokyo) Biotin-labeled DNA was bound to streptoavidin paramagnetic beads (Promega). Approximately 400 μg of UV-irradiated DNA was bound to beads. The cleared lysates were mixed with the UV-DNA affinity beads in 1 ml of binding buffer [salmon sperm DNA (1 mg/ml), 200 mM NaCl, 50 mM tris-HCl (pH 7.4), 1 mM EDTA, and 10% glycerol] After 15 min to allow for binding, the beads were washed twice with 1 ml of washing buffer [350 mM NaCl, 50 mM tris-HCl (pH 7.4), 1 mM EDTA, and 10% glycerol], and protein bound to the beads was eluted with 100 μl of elution buffer [1 M NaCl, 50 mM tris-HCl (pH 7.4), 1 mM EDTA, and 10% glycerol]. After repeating the elution process three times, each 100 μl of eluted solution was combined, and the resultant 300 μl of solution was used as the recombinant Drosophila (6-4)photolyase or control plasmid extract.
13. T Matsunaga et al., Photochem. Photobiol. 57, 934 (1993).
14. A. Yasui et al., EMBO J. 13, 6143 (1994).
15. M. Ahmad and A. R. Cashmore, Nature 366, 162 (1993).
16. K. Malhotra et al , Biochemistry 34, 6892 (1995)
17. S.-T. Kim et al., J Biol. Chem. 269, 8535 (1994).
18. C. Lin et al., Science 269, 968 (1995).
19. H.-W. Park, S.-T Kim, A. Sancar, J. Deisenhofer, ibid 268, 1866 (1995).
20. The recombinant protein migrated as a doublet of ∼62 KD. The lower band may represent a degradation product
21. 2 of UV light and then used in the gel shift assay
22. - strain, as in (7).
23. 2) and purified recombinant (6-4)photolyase were mixed in 400 μl of reaction buffer and exposed to fluorescent light for 30 min. The mixture was phenol extracted, ethanol precipitated, and used for the ELISA assay (7, 70)
24. T. Kato Jr. et al., Nucleic Acids Res. 22, 4119 (1994).
25. K Malhotra, S.-T. Kim, A. Sancar, Biochemistry 33, 8712 (1994)
26. G. D. Small, B. Min, P. A. Lefebrre, Plant Mol Biol. 28, 443 (1995)
27. G J. Barton and M. J E. Sternberg, J Mol Biol 198, 327 (1987).
28. R.M. Schwartz and M. O Dayhoff, in Atlas of Protein Sequence and Structure, M. O. Dayhoff, Ed. (National Biomedical Research Foundation, Washington, DC, 1978), pp. 353-359.
29. We thank H Iwasaki for critical reading of the manuscript. Supported by the Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Science, and Culture of Japan (numbers 05270101 and 07263240) We also thank the Integrated Molecular Analysis of Genome Expression (IMAGE) consortium for the use of their resources.