Evolutionarily diverse LIM domain-containing proteins bind stressed actin filaments through a conserved mechanism.

Jonathan D Winkelman, Caitlin A Anderson, Cristian Suarez, David R Kovar, Margaret L Gardel

Proceedings of the National Academy of Sciences of the United States of America 2020 Oct 13

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The actin cytoskeleton assembles into diverse load-bearing networks, including stress fibers (SFs), muscle sarcomeres, and the cytokinetic ring to both generate and sense mechanical forces. The LIM (Lin11, Isl- 1, and Mec-3) domain family is functionally diverse, but most members can associate with the actin cytoskeleton with apparent force sensitivity. Zyxin rapidly localizes via its LIM domains to failing SFs in cells, known as strain sites, to initiate SF repair and maintain mechanical homeostasis. The mechanism by which these LIM domains associate with stress fiber strain sites (SFSS) is not known. Additionally, it is unknown how widespread strain sensing is within the LIM protein family. We identify that the LIM domain-containing region of 18 proteins from the Zyxin, Paxillin, Tes, and Enigma proteins accumulate to SFSS. Moreover, the LIM domain region from the fission yeast protein paxillin like 1 (Pxl1) also localizes to SFSS in mammalian cells, suggesting that the strain sensing mechanism is ancient and highly conserved. We then used sequence and domain analysis to demonstrate that tandem LIM domains contribute additively, for SFSS localization. Employing in vitro reconstitution, we show that the LIM domain-containing region from mammalian zyxin and fission yeast Pxl1 binds to mechanically stressed F-actin networks but does not associate with relaxed actin filaments. We propose that tandem LIM domains recognize an F-actin conformation that is rare in the relaxed state but is enriched in the presence of mechanical stress.

Citation

Jonathan D Winkelman, Caitlin A Anderson, Cristian Suarez, David R Kovar, Margaret L Gardel. Evolutionarily diverse LIM domain-containing proteins bind stressed actin filaments through a conserved mechanism. Proceedings of the National Academy of Sciences of the United States of America. 2020 Oct 13;117(41):25532-25542