Ullo, M. F. | Search bioRxiv

bioRxiv2021

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

Melanoma cells have been shown to undergo fast amoeboid (leader bleb-based) migration, requiring a single large bleb for migration. In leader blebs, is a rapid flow of cortical actin that drives the cell forward. Using RNAi, we find that co-depleting cofilin-1 and ADF led to a large increase in cortical actin, suggesting that both proteins regulate cortical actin. Furthermore, 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. We found cofilin-1 to be enriched at leader bleb necks, whereas RNAi of cofilin-1 and ADF reduced bleb sizes and the frequency of motile cells. Strikingly, cells without cofilin-1 and ADF had blebs with abnormally long necks. Many of these blebs failed to retract and displayed slow actin turnover. Collectively, our data identifies cofilin-1 and ADF as actin remodeling factors required for fast amoeboid migration.

cell biology↗

bioRxiv2020

Emerin regulation of nuclear stiffness is required for the amoeboid migration of cancer cells in confining environments

When metastasizing, tumor cells must traverse environments with diverse physicochemical properties. Recently, the cell nucleus has emerged as a major regulator of the transition from mesenchymal to fast amoeboid (leader bleb-based) migration. Here, in melanoma cells, we demonstrate that increasing nuclear stiffness through elevating Lamin A, inhibits fast amoeboid migration. Importantly, nuclei may respond to force through stiffening. A key factor in this process is the inner nuclear membrane (INM) protein, emerin. Accordingly, we determined the role of emerin in regulating fast amoeboid migration. Strikingly, we found that both the up- and down-regulation of emerin results in an inhibition of fast amoeboid migration. However, when key Src phosphorylation sites were removed, up-regulation of emerin no longer inhibited fast amoeboid migration. Interestingly, in confined cells, Src activity was low, as measured by a Src biosensor. Thus, the fast amoeboid migration of melanoma cells depends on the precise calibration of emerin activity. Summary StatementIn mechanically constrictive microenvironments, amoeboid migrating melanoma cells require emerin for the precise calibration of nuclear stiffness.

cell biology↗