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Cooperation between mDia1 and ROCK in Rho-induced actin reorganization

Abstract

The small GTPase Rho induces the formation of actin stress fibres and mediates the formation of diverse actin structures. However, it remains unclear how Rho regulates its effectors to elicit such functions. Here we show that GTP-bound Rho activates its effector mDia1 by disrupting mDia1’s intramolecular interactions. Active mDia1 induces the formation of thin actin stress fibres, which are disorganized in the absence of activity of the Rho-associated kinase ROCK. Moreover, active mDia1 transforms ROCK-induced condensed actin fibres into structures reminiscent of Rho-induced stress fibres. Thus mDia1 and ROCK work concurrently during Rho-induced stress-fibre formation. Intriguingly, mDia1 and ROCK, depending on the balance of the two activities, induce actin fibres of various thicknesses and densities. Thus Rho may induce the formation of different actin structures affected by the balance between mDia1 and ROCK signalling.

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Figure 1: F-actin phenotypes of HeLa cells expressing random truncation mutants of mDia1.
Figure 2: Intramolecular interaction in mDia1 and competitive binding of RhoA and the C terminus of mDia1 to the N-terminal region of mDia1.
Figure 3: Phenotypes of HeLa cells expressing mDia1 mutants containing short truncations at the C terminus.
Figure 4: Detailed morphology of actin fibres in cells expressing constitutively active mDia1 and their disorganization under conditions of inactive Rho–ROCK signalling.
Figure 5: F-actin phenotypes of HeLa cells co-transfected with constitutively active mutants of mDia1 and ROCK at various plasmid ratios.
Figure 6: F-actin and focal adhesions induced by active ROCK and mDia1 in HeLa cells expressing the C3 exoenzyme.
Figure 7: Model for activation of mDia1 and two models for actin reorganization downstream of Rho

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Acknowledgements

We thank K. Nonomura for technical assistance; T. Arai and H. Nose for secretarial assistance; and H. A. Popiel for editing the manuscript. N.W. was supported by a JSPS Fellowship in Cancer Research for Young Scientists. This work was supported in part by a Grant-in-Aid for Specially Promoted Research (08102007) from the Ministry of Education, Science, Sports, and Culture of Japan, and by the Human Frontier Science Program.

Correspondence and requests for materials should be addressed to S.N.

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Correspondence to Shuh Narumiya.

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Watanabe, N., Kato, T., Fujita, A. et al. Cooperation between mDia1 and ROCK in Rho-induced actin reorganization . Nat Cell Biol 1, 136–143 (1999). https://doi.org/10.1038/11056

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