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ADF and cofilin-1 collaborate to promote cortical actin flow and the leader bleb-based migration of confined cells

Maria F. Ullo, Jeremy S. Logue
doi: https://doi.org/10.1101/2021.01.11.426289
Maria F. Ullo
Department of Regenerative and Cancer Cell Biology, Albany Medical College, 47 New Scotland Ave, Albany, NY 12208, USA
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Jeremy S. Logue
Department of Regenerative and Cancer Cell Biology, Albany Medical College, 47 New Scotland Ave, Albany, NY 12208, USA
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  • For correspondence: loguej@mail.amc.edu
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Abstract

Melanoma cells have been shown to undergo fast amoeboid or Leader Bleb-Based Migration (LBBM), requiring a large and stable bleb for migration. In leader blebs, is a rapid flow of cortical actin that drives the cell forward. Here, we tested the hypothesis that the actin severing factors, ADF and cofilin-1, are essential for contractility and actin flow for LBBM. Using RNAi in melanoma A375 cells, we find that co-depleting ADF and cofilin-1 led to a large increase in the level of cortical actin, suggesting that ADF and cofilin-1, together, regulate cortical actin in these cells. Moreover, RNAi of ADF and cofilin-1 increased the number of cortical, polymerization competent, barbed-ends. Therefore, severing by these proteins appears to promote cortical actin turnover. Furthermore, actin severing factors can promote contractility through the regulation of actin architecture. However, RNAi of cofilin-1 but not ADF led to a significant decrease in cell stiffness. As LBBM is stimulated by cell confinement, we then used confined cells to evaluate the role of ADF and cofilin-1 in regulating actin dynamics. We found cofilin-1 to be enriched at leader bleb necks, whereas RNAi of ADF and cofilin-1 reduced bleb sizes and the frequency of motile cells. Strikingly, cells without ADF and cofilin-1 had blebs with abnormally long necks. Many blebs failed to retract and displayed slower actin turnover and flow in the absence of ADF and cofilin-1. Collectively, our data identifies ADF and cofilin-1 as actin remodeling factors required for the amoeboid migration of melanoma cells.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted February 22, 2021.
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ADF and cofilin-1 collaborate to promote cortical actin flow and the leader bleb-based migration of confined cells
Maria F. Ullo, Jeremy S. Logue
bioRxiv 2021.01.11.426289; doi: https://doi.org/10.1101/2021.01.11.426289
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ADF and cofilin-1 collaborate to promote cortical actin flow and the leader bleb-based migration of confined cells
Maria F. Ullo, Jeremy S. Logue
bioRxiv 2021.01.11.426289; doi: https://doi.org/10.1101/2021.01.11.426289

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