Drosophila Sec16 mediates the biogenesis of tER sites upstream of Sar1 through an arginine-rich motif.
tER sites are specialized cup-shaped ER subdomains characterized by the focused budding of COPII vesicles. Sec16 has been proposed to be involved in the biogenesis of tER sites by binding to COPII coat components and clustering nascent-coated vesicles. Here, we show that Drosophila Sec16 (dSec16) acts instead as a tER scaffold upstream of the COPII machinery, including Sar1. We show that dSec16 is required for Sar1-GTP concentration to the tER sites where it recruits in turn the components of the COPII machinery to initiate coat assembly. Last, we show that the dSec16 domain required for its localization maps to an arginine-rich motif located in a nonconserved region. We propose a model in which dSec16 binds ER cups via its arginine-rich domain, interacts with Sar1-GTP that is generated on ER membrane by Sec12 and concentrates it in the ER cups where it initiates the formation of COPII vesicles, thus acting as a tER scaffold.
Ivan
Viorica
V
Department of Cell Biology and Institute of Biomembrane, University Medical Centre Utrecht, 3584CX Utrecht, The Netherlands.
de Voer
Gert
G
Xanthakis
Despina
D
Spoorendonk
Kirsten M
KM
Kondylis
Vangelis
V
Rabouille
Catherine
C
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000737:chemistry*; D022181:COP-Coated Vesicles / Q000502:physiology*; D029721:Drosophila Proteins / Q000378:metabolism* / Q000502:physiology; D004331:Drosophila melanogaster / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005786:Gene Expression Regulation; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D020559:Monomeric GTP-Binding Proteins / Q000378:metabolism* / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013347:Subcellular Fractions / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000378:metabolism / Q000502:physiology*
C533275:CG9175 protein, Drosophila; D029721:Drosophila Proteins; D020662:Guanine Nucleotide Exchange Factors; D008565:Membrane Proteins; C096367:SEC16 protein, S cerevisiae; C517555:SEC16A protein, human; D029701:Saccharomyces cerevisiae Proteins; C533274:Sec16 protein, Drosophila; D033921:Vesicular Transport Proteins; D001120:Arginine; C533273:Sar1 protein, Drosophila; D020559:Monomeric GTP-Binding Proteins