Key interactions for clathrin coat stability

Till Böcking; François Aguet; Iris Rapoport; Manuel Banzhaf; Anan Yu; Jean Christophe Zeeh; Tom Kirchhausen

doi:10.1016/j.str.2014.04.002

Key interactions for clathrin coat stability

Till Böcking^*, François Aguet, Iris Rapoport, Manuel Banzhaf, Anan Yu, Jean Christophe Zeeh, Tom Kirchhausen

^*Corresponding author for this work

Molecular Biosciences

Research output: Contribution to journal › Article › peer-review

18 Scopus citations

Abstract

Summary Clathrin-coated vesicles are major carriers of vesicular traffic in eukaryotic cells. This endocytic pathway relies on cycles of clathrin coat assembly and Hsc70-mediated disassembly. Here we identify histidine residues as major determinants of lattice assembly and stability. They are located at the invariant interface between the proximal and distal segments of clathrin heavy chains, in triskelions centered on two adjacent vertices of the coated-vesicle lattice. Mutation of these histidine residues to glutamine alters the pH dependence of coat stability. We then describe single-particle fluorescence imaging experiments in which we follow the effect of these histidine mutations on susceptibility to Hsc70-dependent uncoating. Coats destabilized by these mutations require fewer Hsc70 molecules to initiate disassembly, as predicted by a model in which Hsc70 traps conformational distortions during the auxilin- and Hsc70:ATP-mediated uncoating reaction.

Original language	English (US)
Pages (from-to)	819-829
Number of pages	11
Journal	Structure
Volume	22
Issue number	6
DOIs	https://doi.org/10.1016/j.str.2014.04.002
State	Published - Jun 10 2014

Funding

We thank S.C. Harrison for input into the design of the mutants, discussions, and editorial help; E. Marino for maintaining the Imaging Resource; and members of our laboratories for discussions. T.B. was supported by a fellowship from the Human Frontier Science Program Organization. F.A. was supported by a fellowship from the Swiss National Science Foundation. M.B. was supported by a fellowship from the European Molecular Biology Organization. This work was supported by NIH grants GM-36548 and GM-075252 to T.K. and Australian Research Council grant FT100100411 to T.B.

ASJC Scopus subject areas

Structural Biology
Molecular Biology

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10.1016/j.str.2014.04.002

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title = "Key interactions for clathrin coat stability",

abstract = "Summary Clathrin-coated vesicles are major carriers of vesicular traffic in eukaryotic cells. This endocytic pathway relies on cycles of clathrin coat assembly and Hsc70-mediated disassembly. Here we identify histidine residues as major determinants of lattice assembly and stability. They are located at the invariant interface between the proximal and distal segments of clathrin heavy chains, in triskelions centered on two adjacent vertices of the coated-vesicle lattice. Mutation of these histidine residues to glutamine alters the pH dependence of coat stability. We then describe single-particle fluorescence imaging experiments in which we follow the effect of these histidine mutations on susceptibility to Hsc70-dependent uncoating. Coats destabilized by these mutations require fewer Hsc70 molecules to initiate disassembly, as predicted by a model in which Hsc70 traps conformational distortions during the auxilin- and Hsc70:ATP-mediated uncoating reaction.",

author = "Till B{\"o}cking and Fran{\c c}ois Aguet and Iris Rapoport and Manuel Banzhaf and Anan Yu and Zeeh, {Jean Christophe} and Tom Kirchhausen",

note = "Funding Information: We thank S.C. Harrison for input into the design of the mutants, discussions, and editorial help; E. Marino for maintaining the Imaging Resource; and members of our laboratories for discussions. T.B. was supported by a fellowship from the Human Frontier Science Program Organization. F.A. was supported by a fellowship from the Swiss National Science Foundation. M.B. was supported by a fellowship from the European Molecular Biology Organization. This work was supported by NIH grants GM-36548 and GM-075252 to T.K. and Australian Research Council grant FT100100411 to T.B. ",

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AU - Böcking, Till

AU - Aguet, François

AU - Rapoport, Iris

AU - Banzhaf, Manuel

AU - Yu, Anan

AU - Zeeh, Jean Christophe

AU - Kirchhausen, Tom

N1 - Funding Information: We thank S.C. Harrison for input into the design of the mutants, discussions, and editorial help; E. Marino for maintaining the Imaging Resource; and members of our laboratories for discussions. T.B. was supported by a fellowship from the Human Frontier Science Program Organization. F.A. was supported by a fellowship from the Swiss National Science Foundation. M.B. was supported by a fellowship from the European Molecular Biology Organization. This work was supported by NIH grants GM-36548 and GM-075252 to T.K. and Australian Research Council grant FT100100411 to T.B.

PY - 2014/6/10

Y1 - 2014/6/10

N2 - Summary Clathrin-coated vesicles are major carriers of vesicular traffic in eukaryotic cells. This endocytic pathway relies on cycles of clathrin coat assembly and Hsc70-mediated disassembly. Here we identify histidine residues as major determinants of lattice assembly and stability. They are located at the invariant interface between the proximal and distal segments of clathrin heavy chains, in triskelions centered on two adjacent vertices of the coated-vesicle lattice. Mutation of these histidine residues to glutamine alters the pH dependence of coat stability. We then describe single-particle fluorescence imaging experiments in which we follow the effect of these histidine mutations on susceptibility to Hsc70-dependent uncoating. Coats destabilized by these mutations require fewer Hsc70 molecules to initiate disassembly, as predicted by a model in which Hsc70 traps conformational distortions during the auxilin- and Hsc70:ATP-mediated uncoating reaction.

AB - Summary Clathrin-coated vesicles are major carriers of vesicular traffic in eukaryotic cells. This endocytic pathway relies on cycles of clathrin coat assembly and Hsc70-mediated disassembly. Here we identify histidine residues as major determinants of lattice assembly and stability. They are located at the invariant interface between the proximal and distal segments of clathrin heavy chains, in triskelions centered on two adjacent vertices of the coated-vesicle lattice. Mutation of these histidine residues to glutamine alters the pH dependence of coat stability. We then describe single-particle fluorescence imaging experiments in which we follow the effect of these histidine mutations on susceptibility to Hsc70-dependent uncoating. Coats destabilized by these mutations require fewer Hsc70 molecules to initiate disassembly, as predicted by a model in which Hsc70 traps conformational distortions during the auxilin- and Hsc70:ATP-mediated uncoating reaction.

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Key interactions for clathrin coat stability

Abstract

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