Laboratory of Yeast Genetics and Cell Biology, Rockefeller University, New York, NY 10065, USA.
The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK.
Open Biol. 2023 Mar;13(3):220185. doi: 10.1098/rsob.220185. Epub 2023 Mar 1.
The Rho GTPase family proteins are key regulators of cytoskeletal dynamics. Deregulated activity of Rho GTPases is associated with cancers and neurodegenerative diseases, and their potential as drug targets has long been recognized. Using an economically effective drug screening workflow in fission yeast and human cells, we have identified a Rho GTPase inhibitor, O1. By a suppressor mutant screen in fission yeast, we find a point mutation in the gene that confers resistance to O1. Consistent with the idea that O1 is the direct inhibitor of Rho1, O1 reduced the cellular amount of activated, GTP-bound Rho1 in wild-type cells, but not in the O1-resistant mutant cells, in which the evolutionarily conserved Ala62 residue is mutated to Thr. Similarly, O1 inhibits activity of the human orthologue RhoA GTPase in tissue culture cells. Our studies illustrate the power of yeast phenotypic screens in the identification and characterization of drugs relevant to human cells and have identified a novel GTPase inhibitor for fission yeast and human cells.
Rho GTPase 家族蛋白是细胞骨架动态的关键调节因子。Rho GTPase 的活性失调与癌症和神经退行性疾病有关，其作为药物靶点的潜力早已得到认可。我们使用裂殖酵母和人类细胞中经济有效的药物筛选工作流程，鉴定出一种 Rho GTPase 抑制剂 O1。通过裂殖酵母中的抑制突变体筛选，我们发现了一个基因中的点突变，该突变赋予了对 O1 的抗性。与 O1 是 Rho1 的直接抑制剂的观点一致，O1 减少了野生型细胞中激活的、结合 GTP 的 Rho1 的细胞含量，但在 O1 抗性突变体细胞中没有，在该细胞中，进化上保守的 Ala62 残基突变为 Thr。同样，O1 抑制组织培养细胞中人类同源物 RhoA GTPase 的活性。我们的研究说明了酵母表型筛选在鉴定和表征与人类细胞相关的药物方面的强大功能，并鉴定出一种新型的裂殖酵母和人类细胞的 GTPase 抑制剂。
