Phenotypic analysis of separation-of-function alleles of MEI-41, Drosophila ATM/ATR.
Phenotypic analysis of separation-of-function alleles of MEI-41, Drosophila ATM/ATR.
复制标题
MEI-41、果蝇 ATM/ATR 功能分离等位基因的表型分析。
DOI：
10.1093/genetics/164.2.589
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发表时间：
2003
期刊：
影响因子：
3.3
通讯作者：
Su,TinTin
中科院分区：
文献类型：
--
作者：
Laurençon,Anne;Purdy,Amanda;Sekelsky,Jeff;Hawley,RScott;Su,TinTin
ATM/ATR激酶在真核细胞DNA损伤和复制检查点中作为信号转导子。ATM/ATR同源物的突变具有多效性效应，范围从不育到人类、小鼠和果蝇中基因毒素的杀伤增加。在这里，我们报告了一个无效等位基因ofmei-41，果蝇ATM/ATR同源物的产生，并使用它来记录幼虫有丝分裂检查点和甲基甲磺酸酯（MMS）敏感性的半显性效应。我们还测试了mei-41在最近表征的检查点中的作用，该检查点延迟细胞胚胎中DNA损伤后的中期/后期转换。然后，我们比较了五个现有的mei-41等位基因的空相对于已知的表型（女性不育，细胞周期检查点，和MMS抗性）。我们发现并非所有的表型都受到每个等位基因的同等影响，即，MEI-41在确保生育力、细胞周期调节和对基因毒素的抗性方面的功能在遗传上是可分离的。我们认为MEI-41不是在单一的刚性信号转导途径中起作用，而是在多个分子环境中发挥其许多功能。序列分析确定的突变，其中，大多数等位基因，落在激酶结构域以外的特征不明显的区域;这使我们能够暂时确定MEI-41的其他功能域，可以进行未来的结构-功能研究的这个关键分子。
ATM/ATR kinases act as signal transducers in eukaryotic DNA damage and replication checkpoints. Mutations in ATM/ATR homologs have pleiotropic effects that range from sterility to increased killing by genotoxins in humans, mice, and Drosophila. Here we report the generation of a null allele ofmei-41, Drosophila ATM/ATR homolog, and the use of it to document a semidominant effect on a larval mitotic checkpoint and methyl methanesulfonate (MMS) sensitivity. We also tested the role ofmei-41in a recently characterized checkpoint that delays metaphase/anaphase transition after DNA damage in cellular embryos. We then compare five existingmei-41alleles to the null with respect to known phenotypes (female sterility, cell cycle checkpoints, and MMS resistance). We find that not all phenotypes are affected equally by each allele,i.e., the functions of MEI-41 in ensuring fertility, cell cycle regulation, and resistance to genotoxins are genetically separable. We propose that MEI-41 acts not in a single rigid signal transduction pathway, but in multiple molecular contexts to carry out its many functions. Sequence analysis identified mutations, which, for most alleles, fall in the poorly characterized region outside the kinase domain; this allowed us to tentatively identify additional functional domains of MEI-41 that could be subjected to future structure-function studies of this key molecule.