检查点激酶基因 CHEK2 的功能缺失突变与乳腺癌和其他癌症的风险增加有关。然而，CHEK2 的非同义单核苷酸变异 (SNV) 可能导致 3,188 个独特的氨基酸变化，其中大多数变化对 CHK2 编码蛋白 (CHK2) 功能的影响尚未经过测试。测试变体功能的一种成功方法是测试它们补充 CHEK2 酵母直系同源物 RAD53 突变的能力。这种方法已用于提供超过 100 个 CHEK2 SNV 的功能信息，其结果与人类细胞的功能测定和已知的致病性相一致。在这里，我们使用深度突变扫描 (DMS) 高通量技术测试了 CHEK2 开放阅读框中 4,887 个可能的 SNV 中除两个之外的所有 SNV，以确定它们补充 RAD53 突变体的能力。在非同义变化中，770 个会损害蛋白质功能，而 2,417 个是可以耐受的。结果与先前的结构和功能数据很好地相关，并为临床数据库中确定的所有不确定意义的变异提供了第一个或附加的功能测定。结合起来，这种方法可用于帮助预测在易感性筛查中发现的意义不确定的 CHEK2 变异的致病性，并可应用于其他癌症风险基因。版权所有：© 2024 McCarthy-Leo 等人。这是一篇根据知识共享署名许可条款分发的开放获取文章，允许在任何媒体上不受限制地使用、分发和复制，前提是注明原始作者和来源。

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.Copyright: © 2024 McCarthy-Leo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.