Genome sequencing of large cohorts has identified many protein-coding missense variants, the functional consequences of which remain difficult to predict computationally. Lacoste et al. use a new high-throughput screening approach to characterize the effects of known pathogenic variants on protein localization. They analyzed a total of 3,448 variants, many with known or likely pathogenicity in the ClinVar database, in 1,269 genes by expressing epitope-tagged open reading frames in HeLa cells and characterizing their cellular localization patterns using high-content microscopy. Their screen found that 250 (11%) of the assayable variants (representing 16% of tested genes) resulted in mislocalization compared to the wild-type protein. These variants were enriched for cytoskeletal, Golgi and plasma membrane components; in agreement with this, mislocalization variants often map to transmembrane domains. This microscopy readout could also identify differences in localization of variants associated with a different age-of-onset for disease (PLP1 and GFAP), associated with distinct diseases (ACTB), or observed in cancers (SMAD2). Together, the results from this study demonstrate that the effects of missense variation on protein localization are relatively common and probably relevant to a wide range of disease traits. These and other functional genomics techniques will need to be more widely applied to help clarify the effects of such missense variants.

Original reference: Cell https://doi.org/10.1016/j.cell.2024.09.003 (2024)

大型队列的基因组测序已经确定了许多蛋白质编码错义变异，其功能后果仍然难以通过计算预测。Lacoste 等人使用一种新的高通量筛选方法来表征已知致病性变异对蛋白质定位的影响。他们通过在 HeLa 细胞中表达表位标记的开放阅读框并使用高内涵显微镜表征其细胞定位模式，分析了 1,269 个基因中的总共 3,448 个变异，其中许多在 ClinVar 数据库中具有已知或可能的致病性。他们的筛选发现，与野生型蛋白相比，250 个 （11%） 的可检测变异（占测试基因的 16%）导致定位错误。这些变体富含细胞骨架、高尔基体和质膜成分;与此一致，错误定位的变体通常映射到跨膜结构域。该显微镜读数还可以识别与不同发病年龄 （PLP1 和 GFAP）、与不同疾病相关 （ACTB） 或在癌症中观察到的变异 （SMAD2） 的定位差异。总之，这项研究的结果表明，错义变异对蛋白质定位的影响相对常见，并且可能与广泛的疾病性状有关。这些和其他功能基因组学技术需要更广泛地应用，以帮助阐明此类错义变异的影响。

原始参考： Cell https://doi.org/10.1016/j.cell.2024.09.003 （2024）