Hsc70-induced changes in clathrin-auxilin cage structure suggest a role for clathrin light chains in cage disassembly


The molecular chaperone, Hsc70, together with its cofactor, auxilin, facilitates the ATP-dependent removal of clathrin during clathrin-mediated endocytosis in cells. We have used cryo-electron microscopy to determine the 3D structure of a complex of clathrin, auxilin401-910 and Hsc70 at pH 6 in the presence of ATP, frozen within 20 seconds of adding Hsc70 in order to visualize events that follow the binding of Hsc70 to clathrin and auxilin before clathrin disassembly. In this map,we observe density beneath the vertex of the cage that we attribute to bound Hsc70. This density emerges asymmetrically from the clathrin vertex, suggesting preferential binding by Hsc70 for one of the three possible sites at the vertex. Statistical comparison with a map of whole auxilin and clathrin previously published by us reveals the location of statistically significant differences which implicate involvement of clathrin light chains in structural rearrangements which occur after Hsc70 is recruited. Clathrin disassembly assays using light scattering suggest that loss of clathrin light chains reduces the efficiency with which auxilin facilitates this reaction. These data support a regulatory role for clathrin light chains in clathrin disassembly in addition to their established role in regulating clathrin assembly. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Publication DOI: https://doi.org/10.1111/tra.12085
Divisions: College of Health & Life Sciences > School of Biosciences
College of Health & Life Sciences
College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine
Additional Information: This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. Funding: Royal Society; Medical Research Council (MRC grant) [G120/488, G0601125]; BBSRC; Royal Society Research Grant; NIH grant [GM038093]; Wellcome Trust [055663/Z/98/Z]; NIH [P41 RR001081] Additional Supporting Information may be found in the online version of this article.
Uncontrolled Keywords: 3D structure,cryo-electron microscopy (cryo-EM),endocytosis,molecular chaperone,vesicle uncoating,Biochemistry,Cell Biology,Structural Biology,Molecular Biology,Genetics
Publication ISSN: 1600-0854
Last Modified: 07 Mar 2024 18:06
Date Deposited: 11 Jun 2013 15:00
Full Text Link: http://onlineli ... .12085/abstract
Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2013-09
Published Online Date: 2013-05-25
Authors: Young, Anna
Stoilova-McPhie, Svetla
Rothnie, Alice (ORCID Profile 0000-0002-4259-7015)
Vallis, Yvonne
Harvey-Smith, Phillip
Ranson, Neil
Kent, Helen
Brodsky, Frances M.
Pearse, Barbara M.F.
Roseman, Alan
Smith, Corinne J.



Version: Accepted Version

License: Creative Commons Attribution

Export / Share Citation


Additional statistics for this record