A role for ADP-ribosylation factor in nuclear vesicle dynamics.
Two distinct steps in nuclear envelope assembly can be assayed in vitro: the protein-mediated binding of nuclear-specific vesicles to chromatin, and the subsequent fusion of these vesicles to enclose the chromatin within a double nuclear membrane. Nuclear vesicle fusion, like fusion in the secretory pathway, requires ATP and cytosol and is inhibited by nonhydrolysable GTP analogues. The sensitivity of nuclear vesicle fusion to GTP-gamma S requires a GTP-dependent soluble factor, the properties of which are strikingly similar to a GTP-dependent Golgi binding factor (GGBF) that inhibits Golgi vesicle fusion in the presence of GTP-gamma S and belongs to the ADP-ribosylation factor (ARF) family of small GTPases. In the presence of GTP-gamma S, ARF proteins and alpha-, beta-, gamma-, delta-COP ('coatomer') subunits are associated with Golgi transport vesicles, but the exact roles of ARF proteins in secretion are not yet understood. We report here that purified ARF1 and GGBF have GTP-dependent soluble factor activity in the nuclear vesicle fusion assay. Our results show that the function of ARF is not limited to the Golgi apparatus, and indicate that there may be a link between the formation of nuclear vesicles during mitosis and proteins involved in secretion.
Boman
A L
AL
Department of Cell Biology and Anatomy, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Taylor
T C
TC
Melançon
P
P
Wilson
K L
KL
D020823:ADP-Ribosylation Factor 1; D020727:ADP-Ribosylation Factors; D000818:Animals; D019204:GTP-Binding Proteins / Q000502:physiology*; D006056:Golgi Apparatus / Q000502:physiology; D006160:Guanosine Triphosphate / Q000502:physiology; D066298:In Vitro Techniques; D008561:Membrane Fusion / Q000502:physiology; D009685:Nuclear Envelope / Q000502:physiology*; D011994:Recombinant Proteins; D014981:Xenopus
D011994:Recombinant Proteins; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; D020823:ADP-Ribosylation Factor 1; D020727:ADP-Ribosylation Factors