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Membrane bending begins at any stage of clathrin-coat assembly and defines endocytic dynamics

View ORCID ProfileBrandon L. Scott, Kem A. Sochacki, Shalini T. Low-Nam, Elizabeth M. Bailey, QuocAhn Luu, Amy Hor, Andrea M. Dickey, Steve Smith, Jason G. Kerkvliet, View ORCID ProfileJustin W. Taraska, View ORCID ProfileAdam D. Hoppe
doi: https://doi.org/10.1101/163303
Brandon L. Scott
1Department of Chemistry and Biochemistry, South Dakota State University (SDSU), Brookings, South Dakota, 57007, USA.
2BioSNTR, SDSU
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  • ORCID record for Brandon L. Scott
Kem A. Sochacki
3Laboratory of Molecular Biophysics, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
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Shalini T. Low-Nam
1Department of Chemistry and Biochemistry, South Dakota State University (SDSU), Brookings, South Dakota, 57007, USA.
2BioSNTR, SDSU
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Elizabeth M. Bailey
1Department of Chemistry and Biochemistry, South Dakota State University (SDSU), Brookings, South Dakota, 57007, USA.
2BioSNTR, SDSU
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QuocAhn Luu
4Nanoscience and Nanoengineering, South Dakota School of Mines and Technology (SDSMT), Rapid City, South Dakota, 57701, USA.
5BioSNTR, SDSMT.
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Amy Hor
4Nanoscience and Nanoengineering, South Dakota School of Mines and Technology (SDSMT), Rapid City, South Dakota, 57701, USA.
5BioSNTR, SDSMT.
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Andrea M. Dickey
3Laboratory of Molecular Biophysics, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
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Steve Smith
4Nanoscience and Nanoengineering, South Dakota School of Mines and Technology (SDSMT), Rapid City, South Dakota, 57701, USA.
5BioSNTR, SDSMT.
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Jason G. Kerkvliet
1Department of Chemistry and Biochemistry, South Dakota State University (SDSU), Brookings, South Dakota, 57007, USA.
2BioSNTR, SDSU
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Justin W. Taraska
3Laboratory of Molecular Biophysics, National Heart Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
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  • For correspondence: adam.hoppe@sdstate.edu justin.taraska@nih.gov
Adam D. Hoppe
1Department of Chemistry and Biochemistry, South Dakota State University (SDSU), Brookings, South Dakota, 57007, USA.
2BioSNTR, SDSU
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  • ORCID record for Adam D. Hoppe
  • For correspondence: adam.hoppe@sdstate.edu justin.taraska@nih.gov
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Summary Paragraph

Clathrin-mediated endocytosis internalizes membrane from the cell surface by reshaping flat regions of membrane into spherical vesicles(1, 2). The relationship between membrane bending and clathrin coatomer assembly has been inferred from electron microscopy and structural biology, without directly visualization of membrane bending dynamics (3–6). This has resulted in two distinct and opposing models for how clathrin bends membrane (7–10). Here, polarized Total Internal Reflection Fluorescence microscopy was improved and combined with electron microscopy, atomic force microscopy, and super-resolution imaging to measure membrane bending during endogenous clathrin and dynamin assembly in living cells. Surprisingly, and not predicted by either model, the timing of membrane bending was variable relative to clathrin assembly. Approximately half of the time, membrane bending occurs at the start of clathrin assembly, in the other half, the onset of membrane bending lags clathrin arrival, and occasionally completely assembled flat clathrin transitions into a pit. Importantly, once the membrane bends, the process proceeds to scission with similar timing. We conclude that the pathway of coatomer formation is versatile and can bend the membrane during or after the assembly of the clathrin lattice. These results highlight the heterogeneity in this fundamental biological process, and provide a more complete nanoscale view of membrane bending dynamics during endocytosis.

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Posted July 13, 2017.
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Membrane bending begins at any stage of clathrin-coat assembly and defines endocytic dynamics
Brandon L. Scott, Kem A. Sochacki, Shalini T. Low-Nam, Elizabeth M. Bailey, QuocAhn Luu, Amy Hor, Andrea M. Dickey, Steve Smith, Jason G. Kerkvliet, Justin W. Taraska, Adam D. Hoppe
bioRxiv 163303; doi: https://doi.org/10.1101/163303
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Membrane bending begins at any stage of clathrin-coat assembly and defines endocytic dynamics
Brandon L. Scott, Kem A. Sochacki, Shalini T. Low-Nam, Elizabeth M. Bailey, QuocAhn Luu, Amy Hor, Andrea M. Dickey, Steve Smith, Jason G. Kerkvliet, Justin W. Taraska, Adam D. Hoppe
bioRxiv 163303; doi: https://doi.org/10.1101/163303

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