Formation of stacked ER cisternae by low affinity protein interactions

Erik L. Snapp, Ramanujan S. Hegde, Maura Francolini, Francesca Lombardo, Sara Colombo, Emanuela Pedrazzini, Nica Borgese, Jennifer Lippincott-Schwartz

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313 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)257-269
Number of pages13
JournalJournal of Cell Biology
Volume163
Issue number2
DOIs
StatePublished - Oct 27 2003

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Keywords

  • Cytochrome b5
  • Endoplasmic reticulum
  • FRAP
  • GFP
  • Photobleaching

ASJC Scopus subject areas

  • Cell Biology

Cite this

Snapp, E. L., Hegde, R. S., Francolini, M., Lombardo, F., Colombo, S., Pedrazzini, E., Borgese, N., & Lippincott-Schwartz, J. (2003). Formation of stacked ER cisternae by low affinity protein interactions. Journal of Cell Biology, 163(2), 257-269. https://doi.org/10.1083/jcb.200306020