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

Anna Young, Svetla Stoilova-McPhie, Alice Rothnie, Yvonne Vallis, Phillip Harvey-Smith, Neil Ranson, Helen Kent, Frances M. Brodsky, Barbara M.F. Pearse, Alan Roseman, Corinne J. Smith

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)987-996
Number of pages10
JournalTraffic
Volume14
Issue number9
DOIs
StatePublished - Sep 2013

Keywords

  • 3D structure
  • Cryo-electron microscopy (cryo-EM)
  • Endocytosis
  • Molecular chaperone
  • Vesicle uncoating

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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