Radford Sarah J, Goley Elizabeth, Baxter Kimberly, McMahan Susan, Sekelsky Jeff
Curriculum in Genetics and Molecular Biology, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
Genetics. 2005 Aug;170(4):1737-45. doi: 10.1534/genetics.104.036178. Epub 2005 Jun 8.
Drosophila MEI-9 is the catalytic subunit of a DNA structure-specific endonuclease required for nucleotide excision repair (NER). The enzymatic activity of this endonuclease during NER requires the presence of a second, noncatalytic subunit called ERCC1. In addition to its role in NER, MEI-9 is required for the generation of most meiotic crossovers. To better understand the role of MEI-9 in crossover formation, we report here the characterization of the Drosophila Ercc1 gene. We created an Ercc1 mutant through homologous gene targeting. We find that Ercc1 mutants are identical to mei-9 mutants in sensitivity to DNA-damaging agents, but have a less severe reduction in the number of meiotic crossovers. MEI-9 protein levels are reduced in Ercc1 mutants; however, overexpression of MEI-9 is not sufficient to restore meiotic crossing over in Ercc1 mutants. We conclude that MEI-9 can generate some meiotic crossovers in an ERCC1-independent manner.
果蝇MEI-9是核苷酸切除修复(NER)所需的一种DNA结构特异性核酸内切酶的催化亚基。在NER过程中,这种核酸内切酶的酶活性需要另一种名为ERCC1的非催化亚基的存在。除了在NER中的作用外,MEI-9是大多数减数分裂交叉形成所必需的。为了更好地理解MEI-9在交叉形成中的作用,我们在此报告果蝇Ercc1基因的特征。我们通过同源基因靶向创建了一个Ercc1突变体。我们发现Ercc1突变体在对DNA损伤剂的敏感性方面与mei-9突变体相同,但减数分裂交叉的数量减少程度较轻。Ercc1突变体中MEI-9蛋白水平降低;然而,MEI-9的过表达不足以恢复Ercc1突变体中的减数分裂交叉。我们得出结论,MEI-9可以以不依赖ERCC1的方式产生一些减数分裂交叉。