Light-inducible generation of membrane curvature in live cells with engineered BAR domain proteins

Abstract

Nanoscale membrane curvature is now understood to play an active role in essential cellular processes such as endocytosis, exocytosis and actin dynamics. Previous studies have shown that membrane curvatures directly affect protein functions and intracellular signaling. However, few methods are able to precisely manipulate membrane curvature in live cells. Here, we report the development of a new method of generating nanoscale membrane curvature in live cells that is controllable, reversible, and capable of precise spatial and temporal manipulation. For this purpose, we make use of BAR domain proteins, a family of well-studied membrane-remodeling and membrane-sculpting proteins. Specifically, we engineered two optogenetic systems, opto-FBAR and opto-IBAR, that allow light-inducible formation of positive and negative membrane curvature respectively. Using opto-FBAR, blue light activation results in the formation of tubular membrane invaginations (positive curvature), controllable down to the subcellular level. Using opto-IBAR, blue light illumination results in the formation of membrane protrusions or filopodia (negative curvature). These systems present a novel approach for light-inducible manipulation of nanoscale membrane curvature in live cells. HighlightsO_LIOpto-FBAR enables light-inducible positive membrane curvature formation. C_LIO_LIOpto-IBAR enables light-inducible negative membrane curvature formation. C_LIO_LILight-inducible activation enables precise spatial and temporal control. C_LIO_LIOpto-BAR systems present a new approach for studying membranes in live cells. C_LI