Center for Molecular Medicine and Genetics, Wayne State University School of Medicine, Detroit, Michigan, United States of America.
Department of Oncology, Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, Michigan, United States of America.
PLoS Genet. 2024 Aug 15;20(8):e1011375. doi: 10.1371/journal.pgen.1011375. eCollection 2024 Aug.
Loss of function mutations in the checkpoint kinase gene CHEK2 are associated with increased risk of breast and other cancers. Most of the 3,188 unique amino acid changes that can result from non-synonymous single nucleotide variants (SNVs) of CHEK2, however, have not been tested for their impact on the function of the CHEK2-enocded protein (CHK2). One successful approach to testing the function of variants has been to test for their ability to complement mutations in the yeast ortholog of CHEK2, RAD53. This approach has been used to provide functional information on over 100 CHEK2 SNVs and the results align with functional assays in human cells and known pathogenicity. Here we tested all but two of the 4,887 possible SNVs in the CHEK2 open reading frame for their ability to complement RAD53 mutants using a high throughput technique of deep mutational scanning (DMS). Among the non-synonymous changes, 770 were damaging to protein function while 2,417 were tolerated. The results correlate well with previous structure and function data and provide a first or additional functional assay for all the variants of uncertain significance identified in clinical databases. Combined, this approach can be used to help predict the pathogenicity of CHEK2 variants of uncertain significance that are found in susceptibility screening and could be applied to other cancer risk genes.
失活突变在检查点激酶基因 CHEK2 与增加的风险的乳腺癌和其他癌症。大多数 3188 独特的氨基酸变化，可以从非同义单核苷酸变异 (SNVs) 的 CHEK2，然而，尚未经过测试，对其功能的检查点激酶编码蛋白 (CHK2)。一个成功的方法来测试的功能的变体已测试为他们的能力，以弥补突变酵母同源物的 CHEK2，RAD53。这种方法已被用于提供功能信息超过 100 个 CHEK2 SNVs 的结果与功能测定在人类细胞和已知的致病性。在这里，我们测试了所有但两个 4887 可能的 SNVs 在 CHEK2 开放阅读框为他们的能力，以补充 RAD53 突变体使用高通量技术的深度突变扫描 (DMS)。之间的非同义变化，770 是破坏性的蛋白质功能，而 2417 是容忍。结果与以前的结构和功能数据，并提供了第一个或额外的功能测定所有的变体意义不确定确定在临床数据库。结合起来，这种方法可以用来帮助预测致病性的 CHEK2 变体意义不确定的发现，在易感性筛选，并可应用于其他癌症风险基因。
