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对黑腹果蝇胚胎中紫外线损伤的DNA序列具有高度特异性的新型DNA结合蛋白。

Novel DNA binding proteins highly specific to UV-damaged DNA sequences from embryos of Drosophila melanogaster.

作者信息

Kai M, Takahashi T, Todo T, Sakaguchi K

机构信息

Department of Applied Biological Science, Faculty of Science and Technology, Science University of Tokyo, Chiba-ken, Japan.

出版信息

Nucleic Acids Res. 1995 Jul 25;23(14):2600-7. doi: 10.1093/nar/23.14.2600.

DOI:10.1093/nar/23.14.2600
PMID:7651820
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC307080/
Abstract

Three new proteins which selectively bind to UV-damaged DNA were identified and purified to near homogeneity from UV-irradiated Drosophila melanogaster embryos through several column chromatographies. These proteins, tentatively designated as D-DDB P1, P2 and P3, can be identified as different complex bands in a gel shift assay by using UV-irradiated TC-31 probe DNA. Analysis of the purified D-DDB P1 fraction by native or SDS-polyacrylamide gel electrophoresis and FPLC-Superose 6 gel filtration demonstrated that it is a monomer protein which is a 30 kDa polypeptide. The D-DDB P2 protein is a monopolypeptide with a molecular mass of 14 kDa. Both D-DDB P1 and P2 highly prefer binding to UV-irradiated DNA, and have almost no affinity for non-irradiated DNA. Gel shift assays with either UV-irradiated DNA probes demonstrated that D-DDB P1 may show a preference for binding to (6-4) photoproducts, while D-DDB P2 may prefer binding to pyrimidine dimers. Both these proteins require magnesium ions for binding. D-DDB P1 is an ATP-preferent protein. These findings are discussed in relation to two recently described [Todo and Ryo (1991) Mutat. Res., 273, 85-93; Todo et al. (1993) Nature, 361, 371-374] DNA-binding factors from Drosophila cell extracts. A possible role for these DNA-binding proteins in lesion recognition and DNA-binding proteins in lesion recognition and DNA repair of UV-induced photo-products is discussed.

摘要

通过多次柱色谱法，从紫外线照射的黑腹果蝇胚胎中鉴定并纯化出三种能选择性结合紫外线损伤DNA的新蛋白质，使其纯度接近均一。这些蛋白质暂定为D-DDB P1、P2和P3，在凝胶迁移试验中，使用紫外线照射的TC-31探针DNA可将它们鉴定为不同的复合带。通过天然或SDS-聚丙烯酰胺凝胶电泳以及FPLC-Superose 6凝胶过滤对纯化的D-DDB P1组分进行分析，结果表明它是一种单体蛋白质，为30 kDa的多肽。D-DDB P2蛋白质是一种分子量为14 kDa的单肽。D-DDB P1和P2都高度优先结合紫外线照射的DNA，而对未照射的DNA几乎没有亲和力。用任何一种紫外线照射的DNA探针进行凝胶迁移试验表明，D-DDB P1可能更倾向于结合（6-4）光产物，而D-DDB P2可能更倾向于结合嘧啶二聚体。这两种蛋白质的结合都需要镁离子。D-DDB P1是一种优先结合ATP的蛋白质。结合最近描述的来自果蝇细胞提取物的两种DNA结合因子 [户藤和亮（1991年）《突变研究》，273，85-93；户藤等人（1993年）《自然》，361，371-374] 对这些发现进行了讨论。文中还讨论了这些DNA结合蛋白在紫外线诱导光产物的损伤识别和DNA修复中可能发挥的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05d/307080/2388caaa80b3/nar00014-0044-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05d/307080/04cd491e03d1/nar00014-0041-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05d/307080/ff8f5a375dc3/nar00014-0042-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05d/307080/9a6f0b10108f/nar00014-0043-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05d/307080/9d01b61e21f7/nar00014-0043-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05d/307080/2388caaa80b3/nar00014-0044-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05d/307080/04cd491e03d1/nar00014-0041-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05d/307080/ff8f5a375dc3/nar00014-0042-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05d/307080/9a6f0b10108f/nar00014-0043-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05d/307080/9d01b61e21f7/nar00014-0043-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b05d/307080/2388caaa80b3/nar00014-0044-a.jpg

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