Reconstituting the nuclear envelope and endoplasmic reticulumin vitro

Abstract We describe how in-vitro reconstitution assays are being used to understand the assembly and structure of the nuclear envelope (NE) and endoplasmic reticulum (ER), focussing on membrane dynamics. Our review is partly historical, to honor the cell biologists who began reconstituting nuclear assembly in vitro about 14 years ago. These pioneering complex reconstitution experiments had an enormous impact on our ability to study both organelle assembly and cell cycle regulation. We will illustrate how cell-free exacts from a variety of organisms are now being used to define assembly and disassembly pathways, examine their cell cycle regulation, and test the roles of specific proteins in nuclear and ER formation.