Arf-GEF localization and function at myosin-rich adherens junctions via coiled-coil heterodimerization with an
Shiyu Zheng1, Junior J West1, Cao Guo Yu1
1Department of Cell and Systems Biology, University of Toronto, Toronto, ON M5S 3G5, Canada.
Abstract:
Tissue dynamics require regulated interactions between adherens junctions and cytoskeletal networks. For example, myosin-rich adherens junctions recruit the cytohesin Arf-GEF Steppke, which down-regulates junctional tension and facilitates tissue stretching. We dissected this recruitment mechanism with structure-function and other analyses of Steppke and Stepping stone, an implicated adaptor protein. During Drosophila dorsal closure, Steppke's coiled-coil domain was necessary and sufficient for junctional recruitment. Purified coiled-coil domains of Steppke and Stepping stone heterodimerized through a hydrophobic surface of the Steppke domain. This mapped surface was required for Steppke's junctional localization and tissue regulation. Stepping stone colocalized with Steppke at junctions, and was required for junctional Steppke localization and proper tissue stretching. A second conserved region of Stepping stone was necessary and largely sufficient for junctional localization. Remarkably, this region could substitute for the Steppke coiled-coil domain for junction localization and regulation, suggesting the main role of the Steppke coiled-coil domain is linkage to the junctional targeting region of Stepping stone. Thus, coiled-coil heterodimerization with Stepping stone normally recruits Step to junctions. Intriguingly, Stepping stone's junctional localization also seems partly dependent on Steppke.
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