Superresolution architecture of cornerstone focal adhesions in human pluripotent stem cells

Aki Stubb, Camilo Guzmán, Elisa Närvä, Jesse Aaron, Teng-Leong Chew, Markku Saari, Mitro Miihkinen, Guillaume Jacquemet and Johanna Ivaska ()
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Aki Stubb: University of Turku and Åbo Akademi University
Camilo Guzmán: University of Turku and Åbo Akademi University
Elisa Närvä: University of Turku and Åbo Akademi University
Jesse Aaron: HHMI Janelia Research Campus
Teng-Leong Chew: HHMI Janelia Research Campus
Markku Saari: University of Turku and Åbo Akademi University
Mitro Miihkinen: University of Turku and Åbo Akademi University
Guillaume Jacquemet: University of Turku and Åbo Akademi University
Johanna Ivaska: University of Turku and Åbo Akademi University

Nature Communications, 2019, vol. 10, issue 1, 1-15

Abstract: Abstract While it is clear that key transcriptional programmes are important for maintaining pluripotency, the requirement for cell adhesion to the extracellular matrix remains poorly defined. Human pluripotent stem cells (hPSCs) form colonies encircled by an actin ring and large stable cornerstone focal adhesions (FA). Using superresolution two-colour interferometric photo-activated localisation microscopy, we examine the three-dimensional architecture of cornerstone adhesions and report vertical lamination of FA proteins with three main structural features distinct from previously studied focal adhesions: 1) integrin β5 and talin are present at high density, at the edges of cornerstone FA, adjacent to a vertical kank-rich protein wall, 2) vinculin localises higher than previously reported, displaying a head-above-tail orientation, and 3) surprisingly, actin and α-actinin are present in two discrete z-layers. Finally, we report that depletion of kanks diminishes FA patterning, and actin organisation within the colony, indicating a role for kanks in hPSC colony architecture.

Date: 2019
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DOI: 10.1038/s41467-019-12611-w

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