Jordt Laura Magdalena, Gellert Manuela, Zelms Finja, Bekeschus Sander, Lillig Christopher Horst
The Institute for Medical Biochemistry and Molecular Biology, University Medicine Greifswald, University of Greifswald, Germany.
Clinic and Polyclinic for Dermatology and Venereology University Medicine Rostock, Germany.
FEBS Lett. 2025 May 21;599(16):2334-45. doi: 10.1002/1873-3468.70072.
Glutaredoxin 3 (Grx3) is a multidomain protein (Trx-GrxA-GrxB) with a Trx-like domain and two Grx domains containing a CGFS motif for binding Fe2S2 clusters. To study the function of these domains, HeLa cells with GLRX3 knockout were generated via CRISPR/Cas. The knockout activated iron-regulatory protein 1, indicating iron starvation due to impaired iron metabolism. Transfection with constructs encoding wild-type or individual domains showed that only the Trx-GrxA construct could rescue the phenotype, matching the effect of full-length Grx3. The specific role of the second Grx domain in human Grx3, absent in simpler eukaryotes such as yeast, remains unclear. While the individual domains are insufficient to rescue the knockout of full-length Grx3, the Trx-GrxA module is functionally critical. Impact statement Glutaredoxin 3 (Grx3) contains a Trx-like domain and two Grx domains. The importance of the domains in higher eukaryotes has not previously been addressed in physiological or cellular contexts. Here, we report GLRX3 knockout results in activation of iron regulatory protein 1, and a Trx-GrxA construct could rescue the phenotype.
谷氧还蛋白3（Grx3）是一种多结构域蛋白（Trx-GrxA-GrxB），具有一个类Trx结构域和两个含有用于结合Fe2S2簇的CGFS基序的Grx结构域。为了研究这些结构域的功能，通过CRISPR/Cas技术构建了GLRX3基因敲除的HeLa细胞。基因敲除激活了铁调节蛋白1，表明铁代谢受损导致铁饥饿。用编码野生型或单个结构域的构建体转染后发现，只有Trx-GrxA构建体能够挽救该表型，与全长Grx3的作用效果一致。在酵母等较简单的真核生物中不存在的人类Grx3中第二个Grx结构域的具体作用仍不清楚。虽然单个结构域不足以挽救全长Grx3的基因敲除，但Trx-GrxA模块在功能上至关重要。影响声明：谷氧还蛋白3（Grx3）包含一个类Trx结构域和两个Grx结构域。这些结构域在高等真核生物中的重要性此前在生理或细胞环境中尚未得到研究。在此，我们报告GLRX3基因敲除导致铁调节蛋白1激活，并且Trx-GrxA构建体能够挽救该表型。
