title: Sec12 binds to Sec16 at transitional ER sites.
abstract: COPII vesicles bud from an ER domain known as the transitional ER (tER). Assembly of the COPII coat is initiated by the transmembrane guanine nucleotide exchange factor Sec12. In the budding yeast Pichia pastoris, Sec12 is concentrated at tER sites. Previously, we found that the tER localization of P. pastoris Sec12 requires a saturable binding partner. We now show that this binding partner is Sec16, a peripheral membrane protein that functions in ER export and tER organization. One line of evidence is that overexpression of Sec12 delocalizes Sec12 to the general ER, but simultaneous overexpression of Sec16 retains overexpressed Sec12 at tER sites. Additionally, when P. pastoris Sec12 is expressed in S. cerevisiae, the exogenous Sec12 localizes to the general ER, but when P. pastoris Sec16 is expressed in the same cells, the exogenous Sec12 is recruited to tER sites. In both of these experimental systems, the ability of Sec16 to recruit Sec12 to tER sites is abolished by deleting a C-terminal fragment of Sec16. Biochemical experiments confirm that this C-terminal fragment of Sec16 binds to the cytosolic domain of Sec12. Similarly, we demonstrate that human Sec12 is concentrated at tER sites, likely due to association with a C-terminal fragment of Sec16A. These results suggest that a Sec12-Sec16 interaction has a conserved role in ER export.
authors.affiliation: Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, Illinois, United States of America.
authors.forename: Elisabeth A
authors.initials: EA
authors.lastname: Montegna
authors.forename: Madhura
authors.initials: M
authors.lastname: Bhave
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.forename: Dibyendu
authors.initials: D
authors.lastname: Bhattacharyya
authors.forename: Benjamin S
authors.initials: BS
authors.lastname: Glick
mesh_terms: D001665:Binding Sites; D022181:COP-Coated Vesicles / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D010843:Pichia / Q000737:chemistry; D021381:Protein Transport
chemical_list: D005656:Fungal Proteins; D020662:Guanine Nucleotide Exchange Factors; D008565:Membrane Proteins