Saito Takumi, Ren Yuan, Berro Julien
Department of Molecular Biophysics and Biochemistry, Yale University.
Nanobiology Institute, Yale University.
bioRxiv. 2025 May 14:2025.05.14.653946. doi: 10.1101/2025.05.14.653946.
Cytokinesis is a fundamental process in cell division, where an actomyosin contractile ring plays a central role in completing the cell division. Although some experimental and computational efforts have evaluated ring tension and the molecular organization of rings, the mechanisms of force transmission at the molecular level remain unclear. Here, we used our novel coiled-coil force sensors to measure the force distribution along the formin Cdc12, a key cytokinesis protein in fission yeast. Our force measurements revealed that individual formin Cdc12 molecules transmit up to ~6 pN with distinct mechanisms between the regions upstream and downstream of the formin homology 2 (FH2) domain, which binds the barbed ends of actin filaments. The force transmitted on the N-terminal region before the FH2 domain requires anchoring to a cytokinetic node via the Cdc15 binding region, but is independent of the C-terminal region after FH2. In contrast, the region after the FH2 domain transmits forces independently of the Cdc12's N-terminal region. Force screening using the coiled-coil force sensors found that a region in Cdc12's disordered C-terminal tail can associate with the contractile ring. Altogether, the force measurements with our coiled-coil force sensors allowed us to uncover the mechanisms of force transmission along Cdc12 and characterize new motifs and binding partners.
胞质分裂是细胞分裂中的一个基本过程,其中肌动球蛋白收缩环在完成细胞分裂中起核心作用。尽管一些实验和计算工作已经评估了环张力和环的分子组织,但分子水平上的力传递机制仍不清楚。在这里,我们使用我们新型的卷曲螺旋力传感器来测量沿裂殖酵母中关键胞质分裂蛋白formin Cdc12的力分布。我们的力测量结果表明,单个formin Cdc12分子通过formin同源2(FH2)结构域上游和下游区域之间不同的机制传递高达约6皮牛的力,该结构域结合肌动蛋白丝的尖端。在FH2结构域之前的N端区域传递的力需要通过Cdc15结合区域锚定到胞质分裂节点,但与FH2之后的C端区域无关。相反,FH2结构域之后的区域独立于Cdc12的N端区域传递力。使用卷曲螺旋力传感器进行的力筛选发现,Cdc12无序C端尾部的一个区域可以与收缩环结合。总之,使用我们的卷曲螺旋力传感器进行的力测量使我们能够揭示沿Cdc12的力传递机制,并表征新的基序和结合伙伴。