Zolezzi Francesca, Fuss Jill, Uzawa Satoru, Linn Stuart
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720-3206, USA.
J Biol Chem. 2002 Oct 25;277(43):41183-91. doi: 10.1074/jbc.M207890200. Epub 2002 Aug 13.
Human damaged DNA-binding protein (DDB) is a heterodimer of p48/DDB2 and p127/DDB1 subunits. Mutations in DDB2 are responsible for Xeroderma Pigmentosum group E, but no mutants of mammalian DDB1 have been described. To study DDB1, the Schizosaccharomyces pombe DDB1 sequence homologue (ddb1(+)) was cloned, and a ddb1 deletion strain was constructed. The gene is not essential; however, mutant cells showed a 37% impairment in colony-forming ability, an elongated phenotype, and abnormal nuclei. The ddb1Delta strain was sensitive to UV irradiation, X-rays, methylmethane sulfonate, and thiabendazole, and these sensitivities were compared with those of the well characterized rad13Delta, rhp51Delta, and cds1Delta mutant strains. Ddb1p showed nuclear and nucleolar localization, and the aberrant nuclear structures observed in the ddb1Delta strain suggest a role for Ddb1p in chromosome segregation.
人类损伤DNA结合蛋白(DDB)是由p48/DDB2和p127/DDB1亚基组成的异源二聚体。DDB2中的突变是导致色素性干皮病E组的原因,但尚未描述过哺乳动物DDB1的突变体。为了研究DDB1,克隆了粟酒裂殖酵母DDB1序列同源物(ddb1(+)),并构建了ddb1缺失菌株。该基因并非必需基因;然而,突变细胞的集落形成能力受损37%,呈现伸长的表型且细胞核异常。ddb1Δ菌株对紫外线照射、X射线、甲基磺酸甲酯和噻苯咪唑敏感,并且将这些敏感性与特征明确的rad13Δ、rhp51Δ和cds1Δ突变菌株的敏感性进行了比较。Ddb1p表现出核定位和核仁定位,在ddb1Δ菌株中观察到的异常核结构表明Ddb1p在染色体分离中发挥作用。