Epithelial cells assemble specialized actomyosin structures at E-Cadherin–based cell–cell junctions, and the force exerted drives cell shape change during morphogenesis. The mechanisms that build this supramolecular actomyosin structure remain unclear. We used ZO-knockdown MDCK cells, which assemble a robust, polarized, and highly organized actomyosin cytoskeleton at the zonula adherens, combining genetic and pharmacologic approaches with superresolution microscopy to define molecular machines required. To our surprise, inhibiting individual actin assembly pathways (Arp2/3, formins, or Ena/VASP) did not prevent or delay assembly of this polarized actomyosin structure. Instead, as junctions matured, micron-scale supramolecular myosin arrays assembled, with aligned stacks of myosin filaments adjacent to the apical membrane, overlying disorganized actin filaments. This suggested that myosin arrays might bundle actin at mature junctions. Consistent with this idea, inhibiting ROCK or myosin ATPase disrupted myosin localization/organization and prevented actin bundling and polarization. We obtained similar results in Caco-2 cells. These results suggest a novel role for myosin self-assembly, helping drive actin organization to facilitate cell shape change.
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3 January 2022
Article|
November 23 2021
Micron-scale supramolecular myosin arrays help mediate cytoskeletal assembly at mature adherens junctions
Hui-Chia Yu-Kemp
,
Hui-Chia Yu-Kemp
1
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC
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Rachel A. Szymanski,
Rachel A. Szymanski
2
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC
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Daniel B. Cortes,
Daniel B. Cortes
2
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC
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Nicole C. Gadda
,
Nicole C. Gadda
2
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC
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Madeline L. Lillich
,
Madeline L. Lillich
2
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC
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Amy S. Maddox
,
Amy S. Maddox
1
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC
2
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC
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Mark Peifer
1
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC
2
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC
Correspondence to Mark Peifer: peifer@unc.edu
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Hui-Chia Yu-Kemp
1
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC
Rachel A. Szymanski
2
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC
Daniel B. Cortes
2
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC
Nicole C. Gadda
2
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC
Madeline L. Lillich
2
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC
Amy S. Maddox
1
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC
2
Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC
Correspondence to Mark Peifer: peifer@unc.edu
Received:
March 12 2021
Revision Received:
September 28 2021
Accepted:
October 14 2021
Online Issn: 1540-8140
Print Issn: 0021-9525
Funding
Funder(s):
National Institutes of Health
- Award Id(s): R35 GM118096
Funder(s):
National Institutes of Health
- Award Id(s): T32 CA009156
Funder(s):
National Institutes of Health
- Award Id(s): R01 GM102390
Funder(s):
National Science Foundation
- Award Id(s): 1616661
© 2021 Yu-Kemp et al.
2021
This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).
J Cell Biol (2022) 221 (1): e202103074.
Article history
Received:
March 12 2021
Revision Received:
September 28 2021
Accepted:
October 14 2021
Citation
Hui-Chia Yu-Kemp, Rachel A. Szymanski, Daniel B. Cortes, Nicole C. Gadda, Madeline L. Lillich, Amy S. Maddox, Mark Peifer; Micron-scale supramolecular myosin arrays help mediate cytoskeletal assembly at mature adherens junctions. J Cell Biol 3 January 2022; 221 (1): e202103074. doi: https://doi.org/10.1083/jcb.202103074
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