Abstract
From the fission yeast Schizosaccharomyces pombe, a cDNA fragment was isolated, which confers UV resistance on repair deficient Escherichia coli host cells. The cloned cDNA encodes a protein of 68,815 Da, which has a 36.6% identity of amino acid sequence with the previously identified 74 kDa UV endonuclease of the filamentous fungus Neurospora crassa. Analysis of several truncated gene constructs shows that only the C-terminal two thirds region, which has 54% identity of amino acid sequence with the C-terminal region of the Neurospora homolog, is necessary for complementing activity of UV-sensitivity in the E. coli host cells. Purified recombinant protein from E. coli host cells incises both UV-induced cyclobutane pyrimidine dimers and (6-4) photoproducts at the sites immediately 5' to the DNA damage in the same fashion as the Neurospora protein. Furthermore, a bacterial homologous sequence was isolated from Bacillus subtilis and shows a similar complementing activity of UV sensitivity in E. coli host cells, indicating a wide distribution of this alternative excision repair mechanism in life.
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- Aboussekhra A., Biggerstaff M., Shivji M. K., Vilpo J. A., Moncollin V., Podust V. N., Protić M., Hübscher U., Egly J. M., Wood R. D. Mammalian DNA nucleotide excision repair reconstituted with purified protein components. Cell. 1995 Mar 24;80(6):859–868. doi: 10.1016/0092-8674(95)90289-9. [DOI] [PubMed] [Google Scholar]
- Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
- Birnboim H. C., Nasim A. Excision of pyrimidine dimers by several UV-sensitive mutants of S. pombe. Mol Gen Genet. 1975;136(1):1–8. doi: 10.1007/BF00275444. [DOI] [PubMed] [Google Scholar]
- Bowman K. K., Sidik K., Smith C. A., Taylor J. S., Doetsch P. W., Freyer G. A. A new ATP-independent DNA endonuclease from Schizosaccharomyces pombe that recognizes cyclobutane pyrimidine dimers and 6-4 photoproducts. Nucleic Acids Res. 1994 Aug 11;22(15):3026–3032. doi: 10.1093/nar/22.15.3026. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Freyer G. A., Davey S., Ferrer J. V., Martin A. M., Beach D., Doetsch P. W. An alternative eukaryotic DNA excision repair pathway. Mol Cell Biol. 1995 Aug;15(8):4572–4577. doi: 10.1128/mcb.15.8.4572. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hoeijmakers J. H. Nucleotide excision repair I: from E. coli to yeast. Trends Genet. 1993 May;9(5):173–177. doi: 10.1016/0168-9525(93)90164-d. [DOI] [PubMed] [Google Scholar]
- Malhotra K., Kim S. T., Sancar A. Characterization of a medium wavelength type DNA photolyase: purification and properties of photolyase from Bacillus firmus. Biochemistry. 1994 Jul 26;33(29):8712–8718. doi: 10.1021/bi00195a012. [DOI] [PubMed] [Google Scholar]
- McCready S., Carr A. M., Lehmann A. R. Repair of cyclobutane pyrimidine dimers and 6-4 photoproducts in the fission yeast Schizosaccharomyces pombe. Mol Microbiol. 1993 Nov;10(4):885–890. doi: 10.1111/j.1365-2958.1993.tb00959.x. [DOI] [PubMed] [Google Scholar]
- Quirk P. G., Hicks D. B., Krulwich T. A. Cloning of the cta operon from alkaliphilic Bacillus firmus OF4 and characterization of the pH-regulated cytochrome caa3 oxidase it encodes. J Biol Chem. 1993 Jan 5;268(1):678–685. [PubMed] [Google Scholar]
- Sancar A. Structure and function of DNA photolyase. Biochemistry. 1994 Jan 11;33(1):2–9. doi: 10.1021/bi00167a001. [DOI] [PubMed] [Google Scholar]
- Yajima H., Takao M., Yasuhira S., Zhao J. H., Ishii C., Inoue H., Yasui A. A eukaryotic gene encoding an endonuclease that specifically repairs DNA damaged by ultraviolet light. EMBO J. 1995 May 15;14(10):2393–2399. doi: 10.1002/j.1460-2075.1995.tb07234.x. [DOI] [PMC free article] [PubMed] [Google Scholar]