Parker A E, Van de Weyer I, Laus M C, Verhasselt P, Luyten W H
Department of Experimental Molecular Biology, Janssen Research Foundation, Turnhoutseweg 30, B-2340 Beerse, Belgium.
J Biol Chem. 1998 Jul 17;273(29):18340-6. doi: 10.1074/jbc.273.29.18340.
In the fission yeast Schizosaccharomyces pombe the rad17+ gene is required for both the DNA damage-dependent and the DNA replication-dependent cell cycle checkpoints. We have identified a human cDNA homologue of the S. pombe rad17+ checkpoint gene, designated Hrad17. Hrad17 has 49% identity to the S. pombe rad17+ sequence at the DNA level and 49% identity and 72% similarity at the amino acid level. Northern blot analysis indicates elevated levels of expression in testis and in cancer cell lines. Chromosomal localization by fluorescence in situ hybridization indicates that Hrad17 is located on chromosome 4q13.3-21.2. This region is subject to loss of heterozygosity in several human cancers. To begin to understand the protein-protein interactions of the human checkpoint machinery, we have used the yeast two-hybrid system to examine potential interactions between Hrad1, Hrad9, and Hrad17. We demonstrate a physical interaction between Hrad17 and Hrad1 but no interaction with Hrad9.
在裂殖酵母粟酒裂殖酵母中，rad17+基因对于DNA损伤依赖性和DNA复制依赖性细胞周期检查点都是必需的。我们已经鉴定出粟酒裂殖酵母rad17+检查点基因的一个人类cDNA同源物，命名为Hrad17。Hrad17在DNA水平上与粟酒裂殖酵母rad17+序列有49%的同一性，在氨基酸水平上有49%的同一性和72%的相似性。Northern印迹分析表明在睾丸和癌细胞系中表达水平升高。通过荧光原位杂交进行的染色体定位表明Hrad17位于4号染色体q13.3-21.2区域。该区域在几种人类癌症中会发生杂合性缺失。为了开始了解人类检查点机制的蛋白质-蛋白质相互作用，我们使用酵母双杂交系统来检测Hrad1、Hrad9和Hrad17之间的潜在相互作用。我们证明了Hrad17和Hrad1之间存在物理相互作用，但与Hrad9没有相互作用。
