Disruption of Golgi structure and function in mammalian cells expressing a mutant dynamin

H. Cao, H. M. Thompson, E. W. Krueger, M. A. McNiven

Research output: Contribution to journalArticle

129 Scopus citations

Abstract

The large GTPase dynamin is a mechanoenzyme that participates in the scission of nascent vesicles from the plasma membrane. Recently, dynamin has been demonstrated to associate with the Golgi apparatus in mammalian cells by morphological and biochemical methods. Additional studies using a well characterized, cell-free assay have supported these findings by demonstrating a requirement for dynamin function in the formation of clathrin-coated, and non-clathrin-coated vesicles from the trans-Golgi network (TGN). In this study, we tested if dynamin participates in Golgi function in living cells through the expression of a dominant negative dynamin construct (K44A). Cells co-transfected to express this mutant dynamin and a GFP-tagged Golgi resident protein (TGN38) exhibit Golgi structures that are either compacted, vesiculated, or tubulated. Electron microscopy of these mutant cells revealed large numbers of Golgi stacks comprised of highly tubulated cisternae and an extraordinary number of coated vesicle buds. Cells expressing mutant dynamin and GFP-tagged VSVG demonstrated a marked retention (8- to 11-fold) of the nascent viral G-protein in the Golgi compared to control cells. These observations in living cells are consistent with previous morphological and in vitro studies demonstrating a role for dynamin in the formation of secretory vesicles from the TGN.

Original languageEnglish (US)
Pages (from-to)1993-2002
Number of pages10
JournalJournal of cell science
Volume113
Issue number11
StatePublished - Jun 28 2000

Keywords

  • Dyn 2(aa) K44A
  • Dynamin
  • TGN38
  • VSVG-GFP

ASJC Scopus subject areas

  • Cell Biology

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