Department of Biology, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA 02139, USA; Graduate Program in Biophysics, Harvard University, 86 Brattle Street, Cambridge, MA 02138, USA.
Department of Biophysics, University of Michigan, 1109 Geddes Ave., Ann Arbor, MI 48109, USA.
Curr Biol. 2024 May 20;34(10):2132-2146.e5. doi: 10.1016/j.cub.2024.04.021. Epub 2024 Apr 29.
Actin cortex patterning and dynamics are critical for cell shape changes. These dynamics undergo transitions during development, often accompanying changes in collective cell behavior. Although mechanisms have been established for individual cells' dynamic behaviors, the mechanisms and specific molecules that result in developmental transitions in vivo are still poorly understood. Here, we took advantage of two developmental systems in Drosophila melanogaster to identify conditions that altered cortical patterning and dynamics. We identified a Rho guanine nucleotide exchange factor (RhoGEF) and Rho GTPase activating protein (RhoGAP) pair required for actomyosin waves in egg chambers. Specifically, depletion of the RhoGEF, Ect2, or the RhoGAP, RhoGAP15B, disrupted actomyosin wave induction, and both proteins relocalized from the nucleus to the cortex preceding wave formation. Furthermore, we found that overexpression of a different RhoGEF and RhoGAP pair, RhoGEF2 and Cumberland GAP (C-GAP), resulted in actomyosin waves in the early embryo, during which RhoA activation precedes actomyosin assembly by ∼4 s. We found that C-GAP was recruited to actomyosin waves, and disrupting F-actin polymerization altered the spatial organization of both RhoA signaling and the cytoskeleton in waves. In addition, disrupting F-actin dynamics increased wave period and width, consistent with a possible role for F-actin in promoting delayed negative feedback. Overall, we showed a mechanism involved in inducing actomyosin waves that is essential for oocyte development and is general to other cell types, such as epithelial and syncytial cells.
肌动蛋白皮层的模式形成和动态变化对于细胞形状的改变至关重要。这些动态变化在发育过程中发生转变,通常伴随着细胞集体行为的变化。尽管已经建立了单个细胞动态行为的机制,但对于导致体内发育转变的机制和特定分子仍然知之甚少。在这里,我们利用果蝇中的两个发育系统来确定改变皮层模式形成和动力学的条件。我们鉴定了一个肌动球蛋白波所需的Rho 鸟嘌呤核苷酸交换因子(RhoGEF)和 Rho GTPase 激活蛋白(RhoGAP)对在卵囊中。具体来说,RhoGEF Ect2 或 RhoGAP RhoGAP15B 的耗竭破坏了肌动球蛋白波的诱导,并且这两种蛋白在波形成之前从核重新定位到皮层。此外,我们发现过表达另一种 RhoGEF 和 RhoGAP 对,RhoGEF2 和 Cumberland GAP(C-GAP),导致早期胚胎中的肌动球蛋白波,其中 RhoA 激活在肌动球蛋白组装之前约 4 s 发生。我们发现 C-GAP 被募集到肌动球蛋白波中,并且破坏 F-肌动蛋白聚合改变了波中的 RhoA 信号传导和细胞骨架的空间组织。此外,破坏 F-肌动蛋白动力学增加了波的周期和宽度,这与 F-肌动蛋白在促进延迟负反馈中的可能作用一致。总的来说,我们展示了一种参与诱导肌动球蛋白波的机制,该机制对于卵母细胞发育是必不可少的,并且对其他细胞类型(如上皮细胞和合胞体细胞)也是通用的。