Optogenetic Control of Membrane Tethering and Inter-Membrane Communications

Abstract

Owing to its unsurpassable flexibility and spatiotemporal precision to manipulate biochemical pathways, the field of non-opsin-based optogenetics has been gaining rapid momentum in cell biology over the past decade. By installing light sensitivities into a series of polybasic domains (PB) that mediate phosphoinositide binding in mammalian cells, we report herein the development of a set of single-component optogenetic tools to control protein translocation (termed “optoPB”) toward the plasma membrane, and to further manipulate membrane contact sites between the endoplasmic reticulum (ER) and the plasma membrane (PM). We demonstrated the use of these tools to i) perturb phosphoinositide metabolism and cell signaling on the plasma membrane; ii) reversibly control the formation of ER-PM junctions with high spatial and temporal precision; iii) aid the dissection of molecular determinants that govern protein-phosphoinositide interactions; and iv) photo-tune the distance between ER-PM junctions to restrict protein diffusion on the plasma membrane. These noninvasive optical tools will likely open new possibilities for cell biologists to quantitatively examine the effect of chemical or protein modulators that regulate the dynamics of ER-PM junctions at defined spatiotemporal resolution in living cells. (This work was supported by grants from the NIH (R01GM112003 to Y.Z.) and the Rober A. Welch Foundation (BE-1913 to Y.Z.), the China Scholarship Council (to J.J.) )