TY - JOUR
T1 - Negative membrane curvature catalyzes nucleation of endosomal sorting complex required for transport (ESCRT)-III assembly
AU - Lee, Il Hyung
AU - Kai, Hiroyuki
AU - Carlson, Lars Anders
AU - Groves, Jay T.
AU - Hurley, James H.
N1 - Funding Information:
We thank Peidong Yang and Chong Liu for AFM measurements; Bei Yang, Qingtao Shen, Gerhard Hummer, and Johannes Sch?neberg for advice and discussions; the Berkeley Marvell nanofabrication laboratory and Biomolecular Nanotechnology Center for use of nanofabrication instruments; and the Berkeley CNR Biological Imaging Facility for access to and assistance with SIM microscopy. This work was supported by National Institute of Allergy and Infectious Diseases of the National Institutes of Health Grant R01AI112442. Partial support was provided by the National Cancer Institute of the National Institutes of Health under Award no. U01CA202241(to J.T.G.).
PY - 2015/12/29
Y1 - 2015/12/29
N2 - The endosomal sorting complexes required for transport (ESCRT) machinery functions in HIV-1 budding, cytokinesis, multivesicular body biogenesis, and other pathways, in the course of which it interacts with concave membrane necks and bud rims. To test the role of membrane shape in regulating ESCRT assembly, we nanofabricated templates for invaginated supported lipid bilayers. The assembly of the core ESCRT-III subunit CHMP4B/Snf7 is preferentially nucleated in the resulting 100-nm-deep membrane concavities. ESCRT-II and CHMP6 accelerate CHMP4B assembly by increasing the concentration of nucleation seeds. Superresolution imaging was used to visualize CHMP4B/Snf7 concentration in a negatively curved annulus at the rim of the invagination. Although Snf7 assemblies nucleate slowly on flat membranes, outward growth onto the flat membrane is efficiently nucleated at invaginations. The nucleation behavior provides a biophysical explanation for the timing of ESCRT-III recruitment and membrane scission in HIV-1 budding.
AB - The endosomal sorting complexes required for transport (ESCRT) machinery functions in HIV-1 budding, cytokinesis, multivesicular body biogenesis, and other pathways, in the course of which it interacts with concave membrane necks and bud rims. To test the role of membrane shape in regulating ESCRT assembly, we nanofabricated templates for invaginated supported lipid bilayers. The assembly of the core ESCRT-III subunit CHMP4B/Snf7 is preferentially nucleated in the resulting 100-nm-deep membrane concavities. ESCRT-II and CHMP6 accelerate CHMP4B assembly by increasing the concentration of nucleation seeds. Superresolution imaging was used to visualize CHMP4B/Snf7 concentration in a negatively curved annulus at the rim of the invagination. Although Snf7 assemblies nucleate slowly on flat membranes, outward growth onto the flat membrane is efficiently nucleated at invaginations. The nucleation behavior provides a biophysical explanation for the timing of ESCRT-III recruitment and membrane scission in HIV-1 budding.
KW - HIV-1
KW - Membrane bending
KW - Nanofabrication
KW - Superresolution imaging
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U2 - 10.1073/pnas.1518765113
DO - 10.1073/pnas.1518765113
M3 - Article
C2 - 26668364
AN - SCOPUS:84952690378
VL - 112
SP - 15892
EP - 15897
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 52
ER -
