Calpain-6, a microtubule-stabilizing protein, regulates Rac1 activity and cell motility through interaction with GEF-H1
Abstract
Crosstalk between microtubules and actin filaments is crucial for various cellular functions, including cell migration, spreading and cytokinesis. The Rac1 GTPase plays a key role in such crosstalk at the leading edge of migrating cells in order to promote lamellipodial formation. However, the mechanism underlying the link between microtubules and Rac1 activation remains unclear. Here, we show that calpain-6 (CAPN6), a non-proteolytic calpain with microtubule-binding and -stabilizing activity, might participate in this crosstalk. Small interfering RNA (siRNA)-induced knockdown of Capn6 in NIH 3T3 cells resulted in Rac1 activation, which promoted cell migration, spreading and lamellipodial protrusion. This increase in Rac1 activity was abolished by knockdown of the Rho guanine nucleotide exchange factor GEF-H1 (officially known as Arhgef2). CAPN6 and GEF-H1 colocalized with microtubules and also interacted with each other through specific domains. Upon knockdown of Capn6, GEF-H1 was shown to translocate from microtubules to the lamellipodial region and to interact with Rac1. By contrast, RhoA activity was decreased upon knockdown of Capn6, although low levels of active RhoA or the presence of RhoA molecules appeared to be required for the Capn6-knockdown-induced Rac1 activation. We suggest that CAPN6 acts as a potential regulator of Rac1 activity, through a mechanism involving interaction with GEF-H1, to control lamellipodial formation and cell motility.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Cites
Goll, D. E., Thompson, V. F., Li, H., WEI, W., & Cong, J. (2003) · Physiological Reviews · doi:10.1152/physrev.00029.2002 · Cited by 2,826 · see citing works
Regulation of the small GTP‐binding protein Rho by cell adhesion and the cytoskeleton
Ren, X. D., Kiosses, W. B., & Schwartz, M. A. (1999) · The EMBO Journal · doi:10.1093/emboj/18.3.578 · Cited by 1,573 · see citing works
Coordination of Rho GTPase activities during cell protrusion
Machacek, M., Hodgson, L., Welch, C. M., Elliott, H. L., Pertz, O. C., Nalbant, P., Abell, A. N., Johnson, G. L., Hahn, K. M., & Danuser, G. M. (2009) · Nature · doi:10.1038/nature08242 · Cited by 1,001 · see citing works
Adhesion signaling – crosstalk between integrins, Src and Rho
Huveneers, S., & Danen, E. H. J. (2009) · Journal of Cell Science · doi:10.1242/jcs.039446 · Cited by 845 · see citing works
Conserved microtubule–actin interactions in cell movement and morphogenesis
Rodriguez, O. C., Schaefer, A. W., Mandato, C. A., Forscher, P., Bement, W. M., & Waterman-Storer, C. M. (2003) · Nature Cell Biology · doi:10.1038/ncb0703-599 · Cited by 844 · see citing works
Watanabe, N., Madaule, P., Reid, T., Ishizaki, T., Watanabe, G., Kakizuka, A., Saito, Y., Nakao, K., Jockusch, B. M., & Narumiya, S. (1997) · The EMBO Journal · doi:10.1093/emboj/16.11.3044 · Cited by 823 · see citing works
The many faces of actin: matching assembly factors with cellular structures
Chhabra, E. S., & Higgs, H. N. (2007) · Nature Cell Biology · doi:10.1038/ncb1007-1110 · Cited by 744 · see citing works
Krendel, M., Zenke, F. T., & Bokoch, G. M. (2002) · Nature Cell Biology · doi:10.1038/ncb773 · Cited by 669 · see citing works
mDia mediates Rho-regulated formation and orientation of stable microtubules
Palazzo, A. F., Cook, T. A., Alberts, A. S., & Gundersen, G. G. (2001) · Nature Cell Biology · doi:10.1038/35087035 · Cited by 582 · see citing works
Rho signaling, ROCK and mDia1, in transformation, metastasis and invasion
Narumiya, S., Tanji, M., & Ishizaki, T. (2009) · Cancer and Metastasis Reviews · doi:10.1007/s10555-008-9170-7 · Cited by 530 · see citing works
Microtubule growth activates Rac1 to promote lamellipodial protrusion in fibroblasts
Waterman-Storer, C. M., Worthylake, R. A., Liu, B. P., Burridge, K., & Salmon, E. D. (1999) · Nature Cell Biology · doi:10.1038/9018 · Cited by 491 · see citing works
Beyond polymer polarity: how the cytoskeleton builds a polarized cell
Li, R., & Gundersen, G. G. (2008) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm2522 · Cited by 372 · see citing works
GEF-H1 Couples Nocodazole-induced Microtubule Disassembly to Cell Contractility via RhoA
Chang, Y.-C., Nalbant, P., Birkenfeld, J., Chang, Z.-F., & Bokoch, G. (2008) · Molecular Biology of the Cell · doi:10.1091/mbc.e07-12-1269 · Cited by 357 · see citing works
Calcium-bound structure of calpain and its mechanism of inhibition by calpastatin
Hanna, R. A., Campbell, R. L., & Davies, P. L. (2008) · Nature · doi:10.1038/nature07451 · Cited by 318 · see citing works
C1 domains exposed: From diacylglycerol binding to protein–protein interactions
COLONGONZALEZ, F., & KAZANIETZ, M. (2006) · Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids · doi:10.1016/j.bbalip.2006.05.001 · Cited by 308 · see citing works
Ren, Y., Li, R., Zheng, Y., & Busch, H. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.52.34954 · Cited by 295 · see citing works
Microtubule-induced cortical cell polarityPreprint
Siegrist, S. E., & Doe, C. Q. (2007) · Genes & Development · doi:10.1101/gad.1511207 · Cited by 239 · see citing works
The calpains: modular designs and functional diversity
Croall, D. E., & Ersfeld, K. (2007) · Genome biology · doi:10.1186/gb-2007-8-6-218 · Cited by 221 · see citing works
ROCK and mDia1 antagonize in Rho-dependent Rac activation in Swiss 3T3 fibroblasts
Tsuji, T., Ishizaki, T., Okamoto, M., Higashida, C., Kimura, K., Furuyashiki, T., Arakawa, Y., Birge, R. B., Nakamoto, T., Hirai, H., et al. (2002) · The Journal of Cell Biology · doi:10.1083/jcb.200112107 · Cited by 219 · see citing works
Enomoto, T. (1996) · Cell Structure and Function · doi:10.1247/csf.21.317 · Cited by 218 · see citing works
Benais-Pont, G., Punn, A., Flores-Maldonado, C., Eckert, J. J., Raposo, G., Fleming, T. P., Cereijido, M., Balda, M. S., & Matter, K. (2003) · The Journal of Cell Biology · doi:10.1083/jcb.200211047 · Cited by 202 · see citing works
Localized RhoA Activation as a Requirement for the Induction of Membrane Ruffling
Kurokawa, K., & Matsuda, M. (2005) · Molecular Biology of the Cell · doi:10.1091/mbc.e04-12-1076 · Cited by 193 · see citing works
Birkenfeld, J., Nalbant, P., Yoon, S.-H., & Bokoch, G. M. (2008) · Trends in Cell Biology · doi:10.1016/j.tcb.2008.02.006 · Cited by 190 · see citing works
Zenke, F. T., Krendel, M., DerMardirossian, C. M., King, C. C., Bohl, B. P., & Bokoch, G. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m400084200 · Cited by 181 · see citing works
GEF-H1 is involved in agonist-induced human pulmonary endothelial barrier dysfunction
Birukova, A. A., Adyshev, D., Gorshkov, B., Bokoch, G. M., Birukov, K. G., & Verin, A. D. (2006) · American Journal of Physiology-Lung Cellular and Molecular Physiology · doi:10.1152/ajplung.00259.2005 · Cited by 173 · see citing works
PAK4 mediates morphological changes through the regulation of GEF-H1
Callow, M. G., Zozulya, S., Gishizky, M. L., Jallal, B., & Smeal, T. (2005) · Journal of Cell Science · doi:10.1242/jcs.02313 · Cited by 169 · see citing works
Glaven, J. A., Whitehead, I., Bagrodia, S., Kay, R., & Cerione, R. A. (1999) · Journal of Biological Chemistry · doi:10.1074/jbc.274.4.2279 · Cited by 156 · see citing works
Nalbant, P., Chang, Y.-C., Birkenfeld, J., Chang, Z.-F., & Bokoch, G. M. (2009) · Molecular Biology of the Cell · doi:10.1091/mbc.e09-01-0041 · Cited by 148 · see citing works
Dear, N. T., Matena, K., Vingron, M., & Boehm, T. L. J. (1997) · Genomics · doi:10.1006/geno.1997.4870 · Cited by 136 · see citing works
Barnes, T. M., & Hodgkin, J. (1996) · The EMBO Journal · doi:10.1002/j.1460-2075.1996.tb00825.x · Cited by 108 · see citing works
Ojima, K., Kawabata, Y., Nakao, H., Nakao, K., Doi, N., Kitamura, F., Ono, Y., Hata, S., Suzuki, H., Kawahara, H., et al. (2010) · Journal of Clinical Investigation · doi:10.1172/jci40658 · Cited by 102 · see citing works
Self-citationCalpain 6 Is Involved in Microtubule Stabilization and Cytoskeletal Organization
Tonami, K., Kurihara, Y., Aburatani, H., Uchijima, Y., Asano, T., & Kurihara, H. (2007) · Molecular and Cellular Biology · doi:10.1128/mcb.00992-06 · Cited by 97 · see citing works
Self-citationRhoA/ROCK-mediated switching between Cdc42- and Rac1-dependent protrusion in MTLn3 carcinoma cells
El‐Sibai, M., Pertz, O. C., Pang, H., Yip, S.-C., Lorenz, M., Symons, M. H., Condeelis, J. S., Hahn, K. M., & Backer, J. (2008) · Experimental Cell Research · doi:10.1016/j.yexcr.2008.01.016 · Cited by 95 · see citing works
Hata, S., Abe, M., Suzuki, H., Kitamura, F., Toyama‐Sorimachi, N., Abe, K., Sakimura, K., & Sorimachi, H. (2010) · PLoS Genetics · doi:10.1371/journal.pgen.1001040 · Cited by 82 · see citing works
Self-citationMugita, N., Kimura, Y., Ogawa, M., Saya, H., & Nakao, M. (1997) · Biochemical and Biophysical Research Communications · doi:10.1006/bbrc.1997.7571 · Cited by 36 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Microtubules regulate GEF-H1 in response to extracellular matrix stiffness
Heck, J. N., Ponik, S. M., Garcia-Mendoza, M. G., Pehlke, C. A., Inman, D. R., Eliceiri, K. W., & Keely, P. J. (2012) · Molecular Biology of the Cell · doi:10.1091/mbc.e11-10-0876 · Cited by 96 · see citing works
- Authors
- Tonami, K., Kurihara, Y., Arima, S., Nishiyama, K., Uchijima, Y., Asano, T., Sorimachi, H., & Kurihara, H.
- Affiliations
- Tokyo Metropolitan Institute of Medical Science (JP)
- The University of Tokyo (JP)
- Hiroshima University (JP)
- Venue
- Journal of Cell Science
- Publisher
- The Company of Biologists
- Published
- 2011-04-15
- Volume
- 124
- Issue
- 8
- Pages
- 1214-1223
- ISSN
- 0021-9533 (ISSN-L)
- 1477-9137
- OpenAlex
- W2101579738
- Semantic Scholar
- 108eccd1f266f77dbe7db229434baa450994b28a
- Cited by
- 46
- References
- 35
- Citations by year
- 2026: 1 · 2025: 1 · 2024: 1 · 2023: 3 · 2021: 3 · 2020: 5 · 2019: 3 · 2018: 1 · 2017: 7 · 2016: 3 · 2015: 2 · 2014: 4 · 2013: 5 · 2012: 3
- FWCI
- 2.08
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Cell Science (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Japan Society for the Promotion of Science (23591099)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Calpain Protease Function and Regulation
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
- Keywords
- Rac1, GEF-H1, ARHGEF2, microtubule stabilization, cell motility, lamellipodia, cell spreading, RhoA, Rac1 activation
- MeSH terms
- Animals, Calpain, Cell Movement, Gene Expression Regulation, Microtubules, Protein Binding, Proto-Oncogene Proteins, Pseudopodia, Guanine Nucleotide Exchange Factors, rac1 GTP-Binding Protein, NIH 3T3 Cells, Mice, Rho Guanine Nucleotide Exchange Factors
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review