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A mechano-signalling network linking microtubules, myosin IIA filaments and integrin-based adhesions


The interrelationship between microtubules and the actin cytoskeleton in mechanoregulation of integrin-mediated adhesions is poorly understood. Here, we show that the effects of microtubules on two major types of cell-matrix adhesion, focal adhesions and podosomes, are mediated by KANK family proteins connecting the adhesion protein talin with microtubule tips. Both total microtubule disruption and microtubule uncoupling from adhesions by manipulations with KANKs trigger a massive assembly of myosin IIA filaments, augmenting focal adhesions and disrupting podosomes. Myosin IIA filaments are indispensable effectors in the microtubule-driven regulation of integrin-mediated adhesions. Myosin IIA filament assembly depends on Rho activation by the RhoGEF GEF-H1, which is trapped by microtubules when they are connected with integrin-mediated adhesions via KANK proteins but released after their disconnection. Thus, microtubule capture by integrin-mediated adhesions modulates the GEF-H1-dependent effect of microtubules on the assembly of myosin IIA filaments. Subsequent actomyosin reorganization then remodels the focal adhesions and podosomes, closing the regulatory loop.

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Data availability

All data generated or analysed during this study are included in this published article (and its Supplementary Information files). Raw datasets generated during and/or analysed during the current study are available from the corresponding authors on reasonable request.

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Custom-written code used to analyse the data in the current study is available from the corresponding authors on reasonable request.

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Change history

  • 30 May 2019

    An amendment to this paper has been published and can be accessed via a link at the top of the paper.


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We thank A. Akhmanova (Utrecht University, Netherlands), R. Fäasler (Max Plank Institute for Biochemistry, Martinsried, Germany), C. M. Waterman (National Institutes of Health, USA), R. Zaidel-Bar (Mechanobiology Institute, Singapore) and M. Dodding for providing constructs used in this study. We are grateful to A. Akhmanova for useful discussions and constructive criticism. We thank D. Pitta de Araujo (MBI Science Communications Unit) for help with Fig. 6. This research is supported by the National Research Foundation, Prime Minister’s Office, Singapore, and the Ministry of Education under the Research Centres of Excellence programme through the Mechanobiology Institute, Singapore (ref no. R-714-006-006-271) (A.D.B., N.B.M.R., V.T. and M.N.) and Singapore Ministry of Education Academic Research Fund Tier 3 MOE grant no. MOE2016-T3-1-002 (A.D.B., Y.N.). A.D.B. also acknowledges support from a Maimonides Israeli–France grant (Israeli Ministry of Science Technology and Space) and EU Marie Skłodowska-Curie Network InCeM (project ID 642866) at the Weizmann Institute of Science. N.B.M.R. is also funded by a joint National University of Singapore–King’s College London graduate studentship. G.E.J. is supported by the Medical Research Council, UK (G1100041, MR/K015664) and the generous provision of a visiting professorship from the Mechanobiology Institute, Singapore. P.K. and Z.Z. are funded by the Ministry of Education Academic Research Fund Tier 2 (MOE-T2-1-124), the Mechanobiology Institute seed funding, the National Research Foundation Fellowship (NRF-NRFF-2011-04) and the National Research Foundation Competitive Research Programme (NRF2012NRF-CRP001-084).

Author information

A.D.B. conceived and designed the project together with V.V., P.K. and G.E.J. N.B.M.R. and Y.N. equally designed and performed all experiments and prepared the manuscript; S.V.P., V.T., Z.Z., S.S. and M.N. provided assistance in carrying out experiments and discussed results. A.D.B., N.B.M.R. and Y.N., together with V.V., P.K. and G.E.J., discussed results and prepared the manuscript.

Competing interests

The authors declare no competing interests.

Correspondence to Pakorn Kanchanawong or Gareth E. Jones or Alexander D. Bershadsky.

Supplementary information

Supplementary Information

Supplementary Notes, Supplementary Figs. 1–13, Supplementary Table 1, Supplementary Video Legends 1–17, Supplementary Refs. 1–39

Reporting Summary

Supplementary Video 1

The microtubule plus ends of an HT1080 cell transfected with control siRNA are accumulated at focal adhesions concentrated within adhesive islands

Supplementary Video 2

The microtubule plus ends were randomly distributed over adhesive islands and spaces between them in an HT1080 cell depleted of KANK1 and KANK2

Supplementary Video 3

Microtubule outgrowth induced transient disassembly of focal adhesions in an HT1080 cell

Supplementary Video 4

Rapamycin-induced linking of two parts of the KANK1 molecule triggered disassembly of focal adhesions in an HT1080 cell

Supplementary Video 5

Microtubule disruption triggered massive assembly of myosin II filaments and disassembly of podosomes in a THP-1 cell

Supplementary Video 6

Microtubule disruption triggered massive assembly of myosin II filaments and augmentation of focal adhesion in an HT1080 cell

Supplementary Video 7

Microtubule outgrowth induced disassembly of myosin II filaments in an HT1080 cell

Supplementary Video 8

Transient decrease in traction forces after microtubule outgrowth in an HT1080 cell

Supplementary Video 9

KANK2 knockdown suppressed disassembly of myosin II filaments induced by microtubule outgrowth in an HT1080 cell

Supplementary Video 10

Activation of RhoA by CN03 augmented myosin II filament formation and podosome disruption in a THP-1 cell

Supplementary Video 11

Dynamics of podosomes and myosin II filaments in a control THP-1 cell

Supplementary Video 12

ROCK inhibition by Y-27632 did not affect podosome dynamics in a THP-1 cell

Supplementary Video 13

Inhibition of ROCK by Y-27632 disrupted myosin II filaments and prevented nocodazole-induced disassembly of podosomes in a THP-1 cell

Supplementary Video 14

Disruption of myosin II filaments by Y-27632 resulted in recovery of podosomes in a THP-1 cell pretreated with nocodazole

Supplementary Video 15

Disruption of myosin II filaments by Y-27632 resulted in recovery of podosomes in a KANK1-depleted THP-1 cell

Supplementary Video 16

Constitutively active RhoA and ROCK prevented the disruption of focal adhesions on microtubule outgrowth in HT1080 cells

Supplementary Video 17

GEF-H1 knockdown suppressed disruption of myosin II filaments and prevented disassembly of focal adhesions on microtubule outgrowth in an HT1080 cell

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Fig. 1: Effects of microtubule uncoupling from integrin adhesions by manipulations with KANK proteins.
Fig. 2: KANK proteins are required for targeting microtubules to focal adhesions.
Fig. 3: Myosin II filaments mediate the effect of microtubules on integrin-based adhesions.
Fig. 4: Recovery of functional podosomes in KANK1-depleted cells on ROCK inhibition.
Fig. 5: Function of GEF-H1 in KANK-mediated regulation of integrin-based adhesions.
Fig. 6: Interplay between microtubules, myosin IIA filaments and integrin adhesions.