Multiplexed assay of variant effect reveals residues of functional importance in the BRCA1 coiled-coil and serine cluster domains.
Multiplexed assay of variant effect reveals residues of functional importance in the BRCA1 coiled-coil and serine cluster domains.
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DOI：
10.1371/journal.pone.0293422
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发表时间：
2023
期刊：
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
作者：
从肿瘤抑制蛋白的功能异常变体中区分功能正常变体对于癌症监测、预后和治疗选择至关重要。BRCA1是一种具有许多不确定意义的变体的蛋白质，这些变体尚未被分类为功能正常或异常。当变体尚未通过临床观察分类时，体外功能测定可用于鉴定变体的功能影响。在这里，我们采用同源性定向修复（HDR）报告分析，以评估超过300个错义和无义BRCA1的氨基酸残基1280和1576之间的变体，其中包括卷曲螺旋和丝氨酸簇结构域。功能异常的变体倾向于聚集在已知与PALB 2相互作用的残基中，这对于同源性定向修复至关重要。通过单次试验和ClinVar数据库变体判读确认多重结果。多重检测结果与ClinVar数据库中指定的良性或可能良性或致病或可能致病变体的比较，多重检测的特异性和灵敏度均为100%。临床医生可以参考这种功能测定的结果，以帮助指导癌症治疗和监测选择。这些结果是第一次使用多重方法评估BRCA1的该结构域，并表明该结构域在DNA修复过程中的重要性。
Delineating functionally normal variants from functionally abnormal variants in tumor suppressor proteins is critical for cancer surveillance, prognosis, and treatment options. BRCA1 is a protein that has many variants of uncertain significance which are not yet classified as functionally normal or abnormal. In vitro functional assays can be used to identify the functional impact of a variant when the variant has not yet been categorized through clinical observation. Here we employ a homology-directed repair (HDR) reporter assay to evaluate over 300 missense and nonsense BRCA1 variants between amino acid residues 1280 and 1576, which encompasses the coiled-coil and serine cluster domains. Functionally abnormal variants tended to cluster in residues known to interact with PALB2, which is critical for homology-directed repair. Multiplexed results were confirmed by singleton assay and by ClinVar database variant interpretations. Comparison of multiplexed results to designated benign or likely benign or pathogenic or likely pathogenic variants in the ClinVar database yielded 100% specificity and 100% sensitivity of the multiplexed assay. Clinicians can reference the results of this functional assay for help in guiding cancer treatment and surveillance options. These results are the first to evaluate this domain of BRCA1 using a multiplexed approach and indicate the importance of this domain in the DNA repair process.
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影响因子：
7.3
作者：
Kettenbach AN;Schweppe DK;Faherty BK;Pechenick D;Pletnev AA;Gerber SA
通讯作者：
Gerber SA
影响因子：
64.8
作者：
Findlay GM;Daza RM;Martin B;Zhang MD;Leith AP;Gasperini M;Janizek JD;Huang X;Starita LM;Shendure J
通讯作者：
Shendure J
影响因子：
4.5
作者：
通讯作者：
--
影响因子：
6
作者：
Clark, Serena L.;Rodriguez, Ana M.;Boehning, Darren
通讯作者：
Boehning, Darren
影响因子：
--
作者：
Beckta, Jason M.;Dever, Seth M.;Valerie, Kristoffer
通讯作者：
Valerie, Kristoffer