Formation of Stacked ER Cisternae by Low Affinity Protein Interactions

• Published in: The Journal of cell biology Vol. 163; no. 2; pp. 257 - 269
• Main Authors: Snapp, Erik L., Hedge, Ramanujan S., Francolini, Maura, Lombardo, Francesca, Colombo, Sara, Pedrazzini, Emanuela, Borgese, Nica, Lippincott-Schwartz, Jennifer
• Format: Journal Article
• Language: English
• Published: United States Rockefeller University Press 27.10.2003; The Rockefeller University Press
• Subjects: Animals; Cell Line; Cell membranes; Cells; Cellular biology; Cercopithecus aethiops; Chimeras; COS Cells; Endoplasmic reticulum; Endoplasmic Reticulum - metabolism; Endoplasmic Reticulum - ultrastructure; Endoplasmic Reticulum, Smooth - metabolism; Endoplasmic Reticulum, Smooth - ultrastructure; Epithelial cells; Fluorescence; Green Fluorescent Proteins; Hepatocytes; Intracellular Membranes - metabolism; Intracellular Membranes - ultrastructure; Kidney cells; Light; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Membrane Proteins - chemistry; Membrane Proteins - metabolism; Membranes; Models, Biological; Neurons; Point Mutation; Protein Structure, Tertiary; Proteins; Recombinant Fusion Proteins - chemistry; Recombinant Fusion Proteins - metabolism
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.200306020

The endoplasmic reticulum (ER) can transform from a network of branching tubules into stacked membrane arrays (termed organized smooth ER [OSER]) in response to elevated levels of specific resident proteins, such as cytochrome b(5). Here, we have tagged OSER-inducing proteins with green fluorescent protein (GFP) to study OSER biogenesis and dynamics in living cells. Overexpression of these proteins induced formation of karmellae, whorls, and crystalloid OSER structures. Photobleaching experiments revealed that OSER-inducing proteins were highly mobile within OSER structures and could exchange between OSER structures and surrounding reticular ER. This indicated that binding interactions between proteins on apposing stacked membranes of OSER structures were not of high affinity. Addition of GFP, which undergoes low affinity, antiparallel dimerization, to the cytoplasmic domains of non-OSER-inducing resident ER proteins was sufficient to induce OSER structures when overexpressed, but addition of a nondimerizing GFP variant was not. These results point to a molecular mechanism for OSER biogenesis that involves weak homotypic interactions between cytoplasmic domains of proteins. This mechanism may underlie the formation of other stacked membrane structures within cells.