TY - JOUR
T1 - Solution structure of the Atg1 complex
T2 - Implications for the architecture of the phagophore assembly site
AU - Köfinger, Jürgen
AU - Ragusa, Michael J.
AU - Lee, Il Hyung
AU - Hummer, Gerhard
AU - Hurley, James H.
N1 - Funding Information:
We thank Greg Hura for help with SAXS data collection and interpretation and Jason Brickner and Will Prinz for yeast strains. This work was supported by the NIH GM111730 (J.H.H), Ruth Kirschtein NRSA Fellowship GM099319 (M.J.R.), and the Max Planck Society (J.K., G.H.). This work was conducted in part at the Advanced Light Source (ALS), a national user facility operated by Lawrence Berkeley National Laboratory on behalf of the Department of Energy, Office of Basic Energy Sciences, through the Integrated Diffraction Analysis Technologies (IDAT) program, supported by DOE Office of Biological and Environmental Research and NIH grant GM105404 .
Publisher Copyright:
© 2015 Elsevier Ltd.
PY - 2015/5/5
Y1 - 2015/5/5
N2 - The biogenesis of autophagosomes commences at the phagophore assembly site (PAS), a protein-vesicle ultrastructure that is organized by the Atg1 complex. The Atg1 complex consists of the Atg1 protein kinase, the intrinsically disordered region-rich Atg13, and the dimeric double crescent-shaped Atg17-Atg31-Atg29 subcomplex. We show that the PAS contains a relatively uniform ∼28 copies of Atg17, and upon autophagy induction, similar numbers of Atg1 and Atg13 molecules. We then apply ensemble refinement of small-angle X-ray scattering to determine the solution structures of the Atg1-Atg13 and Atg17-Atg31-Atg29 subcomplexes and the Atg1 complex, using a trimmed minipentamer tractable to biophysical studies. We observe tetramers of Atg1 pentamers that assemble via Atg17-Atg31-Atg29. This leads to a model for the higher organization of the Atg1 complex in PAS scaffolding.
AB - The biogenesis of autophagosomes commences at the phagophore assembly site (PAS), a protein-vesicle ultrastructure that is organized by the Atg1 complex. The Atg1 complex consists of the Atg1 protein kinase, the intrinsically disordered region-rich Atg13, and the dimeric double crescent-shaped Atg17-Atg31-Atg29 subcomplex. We show that the PAS contains a relatively uniform ∼28 copies of Atg17, and upon autophagy induction, similar numbers of Atg1 and Atg13 molecules. We then apply ensemble refinement of small-angle X-ray scattering to determine the solution structures of the Atg1-Atg13 and Atg17-Atg31-Atg29 subcomplexes and the Atg1 complex, using a trimmed minipentamer tractable to biophysical studies. We observe tetramers of Atg1 pentamers that assemble via Atg17-Atg31-Atg29. This leads to a model for the higher organization of the Atg1 complex in PAS scaffolding.
UR - http://www.scopus.com/inward/record.url?scp=84930188743&partnerID=8YFLogxK
U2 - 10.1016/j.str.2015.02.012
DO - 10.1016/j.str.2015.02.012
M3 - Article
C2 - 25817386
AN - SCOPUS:84930188743
SN - 0969-2126
VL - 23
SP - 809
EP - 818
JO - Structure
JF - Structure
IS - 5
ER -