Sec16 influences transitional ER sites by regulating rather than organizing COPII. Author Nike Bharucha, Yang Liu, Effrosyni Papanikou, Conor McMahon, Masatoshi Esaki, Philip Jeffrey, Frederick Hughson, Benjamin Glick Publication Year 2013 Type Journal Article Abstract During the budding of coat protein complex II (COPII) vesicles from transitional endoplasmic reticulum (tER) sites, Sec16 has been proposed to play two distinct roles: negatively regulating COPII turnover and organizing COPII assembly at tER sites. We tested these ideas using the yeast Pichia pastoris. Redistribution of Sec16 to the cytosol accelerates tER dynamics, supporting a negative regulatory role for Sec16. To evaluate a possible COPII organization role, we dissected the functional regions of Sec16. The central conserved domain, which had been implicated in coordinating COPII assembly, is actually dispensable for normal tER structure. An upstream conserved region (UCR) localizes Sec16 to tER sites. The UCR binds COPII components, and removal of COPII from tER sites also removes Sec16, indicating that COPII recruits Sec16 rather than the other way around. We propose that Sec16 does not in fact organize COPII. Instead, regulation of COPII turnover can account for the influence of Sec16 on tER sites. Keywords Binding Sites, Protein Binding, Mutation, Microscopy, Fluorescence, Green Fluorescent Proteins, Immunoblotting, Two-Hybrid System Techniques, Endoplasmic Reticulum, Protein Transport, Cytosol, COP-Coated Vesicles, Fungal Proteins, Pichia Journal Mol Biol Cell Volume 24 Issue 21 Pages 3406-19 Date Published 2013 Nov ISSN Number 1939-4586 DOI 10.1091/mbc.E13-04-0185 Alternate Journal Mol Biol Cell PMCID PMC3814151 PMID 24006484 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML