title: Dynamic nucleation of Golgi apparatus assembly from the endoplasmic reticulum in interphase hela cells.
abstract: Models of Golgi apparatus biogenesis and maintenance are focused on two possibilities: one is self-assembly from the endoplasmic reticulum, and the other is nucleation by a stable template. Here, we asked in three different experimental situations whether assembly of the Golgi apparatus might be dynamically nucleated. During microtubule depolymerization, the integral membrane protein p27 and the peripheral Golgi protein GM130, appeared in newly formed, scattered Golgi elements before three different Golgi apparatus cisternal enzymes, whereas GRASP55, a medial peripheral Golgi protein, showed, if anything, a tendency to accumulate in scattered Golgi elements later than a cisternal enzyme. During Golgi formation after brefeldin A washout, endoplasmic reticulum exit of Golgi resident enzymes could be completely separated from that of p27 and GM130. p27 and GM130 accumulation was onto newly organized perinuclear structures, not brefeldin A remnants, and preceded that of a cisternal enzyme. Reassembly was completely sensitive to guanosine 5'-diphosphate-restricted Sar1p. When cells were microinjected with Sar1pWT DNA to reverse a guanosine 5'-diphosphate-restricted Sar1p endoplasmic reticulum-exit block phenotype, GM130 and p27 collected perinuclearly with little to no exit of a cisternal enzyme from the endoplasmic reticulum. The overall data strongly indicate that the assembly of the Golgi apparatus can be nucleated dynamically by GM130/p27 associated structures. We define dynamic nucleation as the first step in a staged organelle assembly process in which new component association forms a microscopically visible structure onto which other components add later, e.g. Golgi cisternae.
authors.affiliation: Department of Physiology and Biophysics, University of Arkansas for Medical Sciences (UAMS), 4301 West Markham Street, Little Rock, AR 72205, USA.
authors.forename: Murat
authors.initials: M
authors.lastname: Kasap
authors.forename: Stephanie
authors.initials: S
authors.lastname: Thomas
authors.forename: Erin
authors.initials: E
authors.lastname: Danaher
authors.forename: Virginia
authors.initials: V
authors.lastname: Holton
authors.forename: Shu
authors.initials: S
authors.lastname: Jiang
authors.forename: Brian
authors.initials: B
authors.lastname: Storrie
mesh_terms: D000970:Antineoplastic Agents / Q000378:metabolism; D001324:Autoantigens; D015415:Biomarkers; D020126:Brefeldin A / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism* / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D006367:HeLa Cells / Q000166:cytology*; D006801:Humans; D007399:Interphase / Q000502:physiology*; D008565:Membrane Proteins / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D020559:Monomeric GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism; D017350:N-Acetylgalactosaminyltransferases / Q000378:metabolism; D015739:Nocodazole / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000378:metabolism; D000097763:Polypeptide N-acetylgalactosaminyltransferase
chemical_list: D000970:Antineoplastic Agents; D001324:Autoantigens; D015415:Biomarkers; D020126:Brefeldin A; D008565:Membrane Proteins; D020559:Monomeric GTP-Binding Proteins; D017350:N-Acetylgalactosaminyltransferases; D015739:Nocodazole; D011500:Protein Synthesis Inhibitors; D000097763:Polypeptide N-acetylgalactosaminyltransferase; C097741:Golgin subfamily A member 2; C493083:SAR1A protein, human; C513282:SAR1B protein, human