Gebbia Marinella, Zimmerman Daniel, Jiang Rosanna, Nguyen Maria, Weile Jochen, Li Roujia, Gavac Michelle, Kishore Nishka, Sun Song, Boonen Rick A, Hamilton Rayna, Dines Jennifer N, Wahl Alexander, Reuter Jason, Johnson Britt, Fowler Douglas M, Couch Fergus J, van Attikum Haico, Roth Frederick P
The Donnelly Centre, University of Toronto, Toronto, ON, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada; Lunenfeld-Tanenbaum Research Institute, Sinai Health, Toronto, ON, Canada.
Leiden University Medical Center, Leiden, the Netherlands.
Am J Hum Genet. 2024 Dec 5;111(12):2675-2692. doi: 10.1016/j.ajhg.2024.10.013.
The tumor suppressor CHEK2 encodes the serine/threonine protein kinase CHK2 which, upon DNA damage, is important for pausing the cell cycle, initiating DNA repair, and inducing apoptosis. CHK2 phosphorylation of the tumor suppressor BRCA1 is also important for mitotic spindle assembly and chromosomal stability. Consistent with its cell-cycle checkpoint role, both germline and somatic variants in CHEK2 have been linked to breast and other cancers. Over 90% of clinical germline CHEK2 missense variants are classified as variants of uncertain significance, complicating diagnosis of CHK2-dependent cancer. We therefore sought to test the functional impact of all possible missense variants in CHK2. Using a scalable multiplexed assay based on the ability of human CHK2 to complement DNA sensitivity of Saccharomyces cerevisiae cells lacking the CHEK2 ortholog, RAD53, we generated a systematic "missense variant effect map" for CHEK2 missense variation. The map reflects known biochemical features of CHK2 while offering new biological insights. It also provides strong evidence toward pathogenicity for some clinical missense variants and supporting evidence toward benignity for others. Overall, this comprehensive missense variant effect map contributes to understanding of both known and yet-to-be-observed CHK2 variants.
肿瘤抑制基因CHEK2编码丝氨酸/苏氨酸蛋白激酶CHK2，在DNA损伤时，CHK2对于暂停细胞周期、启动DNA修复以及诱导细胞凋亡至关重要。肿瘤抑制基因BRCA1的CHK2磷酸化对于有丝分裂纺锤体组装和染色体稳定性也很重要。与其细胞周期检查点作用一致，CHEK2中的种系和体细胞变异均与乳腺癌及其他癌症有关。超过90%的临床种系CHEK2错义变异被归类为意义未明的变异，这使得CHK2依赖性癌症的诊断变得复杂。因此，我们试图测试CHK2中所有可能的错义变异的功能影响。利用基于人类CHK2能够补充缺乏CHK2直系同源基因RAD53的酿酒酵母细胞的DNA敏感性的可扩展多重检测方法，我们生成了CHK2错义变异的系统“错义变异效应图谱”。该图谱反映了CHK2已知的生化特征，同时提供了新的生物学见解。它还为一些临床错义变异的致病性提供了有力证据，为其他变异的良性提供了支持性证据。总体而言，这一全面的错义变异效应图谱有助于理解已知的和尚未观察到的CHK2变异。
