Sar1 assembly regulates membrane constriction and ER export.
The guanosine triphosphatase Sar1 controls the assembly and fission of COPII vesicles. Sar1 utilizes an amphipathic N-terminal helix as a wedge that inserts into outer membrane leaflets to induce vesicle neck constriction and control fission. We hypothesize that Sar1 organizes on membranes to control constriction as observed with fission proteins like dynamin. Sar1 activation led to membrane-dependent oligomerization that transformed giant unilamellar vesicles into small vesicles connected through highly constricted necks. In contrast, membrane tension provided through membrane attachment led to organization of Sar1 in ordered scaffolds that formed rigid, uniformly nonconstricted lipid tubules to suggest that Sar1 organization regulates membrane constriction. Sar1 organization required conserved residues located on a unique C-terminal loop. Mutations in this loop did not affect Sar1 activation or COPII recruitment and enhanced membrane constriction, yet inhibited Sar1 organization and procollagen transport from the endoplasmic reticulum (ER). Sar1 activity was directed to liquid-disordered lipid phases. Thus, lipid-directed and tether-assisted Sar1 organization controls membrane constriction to regulate ER export.
Long
Kimberly R
KR
Department of Cell Biology and Physiology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USA.
Yamamoto
Yasunori
Y
Baker
Adam L
AL
Watkins
Simon C
SC
Coyne
Carolyn B
CB
Conway
James F
JF
Aridor
Meir
M
D000818:Animals; D001692:Biological Transport / Q000502:physiology; D022181:COP-Coated Vesicles / Q000235:genetics / Q000378:metabolism*; D018938:Caco-2 Cells; D002462:Cell Membrane / Q000235:genetics / Q000378:metabolism*; D004285:Dogs; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology; D006801:Humans; D020559:Monomeric GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D055503:Protein Multimerization / Q000502:physiology; D017433:Protein Structure, Secondary
C493083:SAR1A protein, human; D020559:Monomeric GTP-Binding Proteins