用于分析与严重先天性中性粒细胞减少症相关的SRP54突变功能的酵母模型
A yeast model for the functional analysis of SRP54 mutations associated with severe congenital neutropenia.
摘要(中文译文)
人类SRP54基因的突变与严重先天性中性粒细胞减少症（SCN）的病理生理有关。SRP54是一种关键蛋白，是信号识别颗粒六个蛋白亚基之一，负责将蛋白质共翻译靶向到内质网。几种SRP54突变体的晶体结构和生化表征为SRP54突变如何影响其功能提供了见解。然而，目前尚无可扩展、灵活的平台来研究SRP54突变的序列-结构-功能关系。在此，我们构建了一株酿酒酵母菌株，其生长依赖于激素诱导的SRP54表达。我们证明了该模型适用于研究SRP54以及与直系同源SRP54突变体相关的疑似显性负性表型。最常见的酵母直系同源突变体S125del（人直系同源T117del）表现出最不严重的生长缺陷；相反，G234E突变体（人直系同源G226E）表现出最严重的生长缺陷，这与SCN的临床表现一致。最后，我们对临床数据中发现的四个此前未表征的SRP54突变进行了表征。我们还测定了酿酒酵母SRP54的首个X射线结构。野生型蛋白的NG结构域显示出与来自各门的同源物预测的高度结构相似性。该结构揭示了酵母蛋白的独特特征，并为突变表型提供了结构解释。该单倍体模型能够在适合高通量筛选方法的同时重现这些表型，使其成为研究SRP54的有力工具。此外，用于创建该模型的方法也可用于研究其他涉及必需基因和准必需基因的人类疾病。
Abstract
Mutations in the human SRP54 gene are linked to the pathophysiology of severe congenital neutropenia (SCN). SRP54 is a key protein comprising one of the six protein subunits of the signal recognition particle responsible for cotranslational targeting of proteins to the ER. Crystal structures and biochemical characterization of several SRP54 mutants provide insights into how SRP54 mutations affect its function. However, no scalable, flexible platform exists to study the sequence-structure-function relationships of SRP54 mutations. Here, we engineered a Saccharomyces cerevisiae strain whose growth depends on hormone-inducible SRP54 expression. We demonstrate the model's suitability for studying SRP54 and the suspected dominant-negative phenotypes associated with orthologous SRP54 mutants. The most common yeast orthologue, S125del (T117del human orthologue), displayed the least severe growth defect; in contrast, the G234E mutant (G226E human orthologue) displayed the most severe growth defect, in line with clinical presentations in SCN. Lastly, we characterized four previously uncharacterized SRP54 mutations found in clinical data. We also determined the first X-ray structure of Saccharomyces cerevisiae SRP54. The NG domain of the wild-type protein shows predicted high structural similarity to homologs from across phyla. The structure outlines unique features of the yeast protein, along with providing a structural explanation for mutant phenotypes. The ability of this haploid model to recapitulate these phenotypes while remaining amenable to high-throughput screening approaches makes it a powerful tool for studying SRP54. Furthermore, the methodology used to create this model may also be used to study other human diseases involving essential and quasi-essential genes.
如何引用
AMA
Lawton F Long, Shivani Biskunda, Ming Yang, Tiffany Zhou, Grace Schlichting, Ava Domson, et al. A yeast model for the functional analysis of SRP54 mutations associated with severe congenital neutropenia.. The FEBS journal. 2026; doi:10.1111/febs.70725.
APA
Lawton F Long, Shivani Biskunda, Ming Yang, Tiffany Zhou, Grace Schlichting, Ava Domson, et al (2026). A yeast model for the functional analysis of SRP54 mutations associated with severe congenital neutropenia.. The FEBS journal. https://doi.org/10.1111/febs.70725
GB/T 7714
Lawton F Long, Shivani Biskunda, Ming Yang, Tiffany Zhou, Grace Schlichting, Ava Domson, et al. A yeast model for the functional analysis of SRP54 mutations associated with severe congenital neutropenia.[J]. The FEBS journal, 2026 doi:10.1111/febs.70725.
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