Previous Article | Next Article 
Molecular and Cellular Biology, December 2001, p. 8022-8034, Vol. 21, No. 23
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.23.8022-8034.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Kinectin Is a Key Effector of RhoG
Microtubule-Dependent Cellular Activity
E.
Vignal,1
A.
Blangy,1
M.
Martin,2
C.
Gauthier-Rouvière,1 and
P.
Fort1,*
Centre de Recherche en Biochimie
Macromoléculaire, CNRS-UPR1086, 34293 Montpellier cedex
5,1 and Laboratoire de Dynamique
Cellulaire, CNRS-UMR5539, Université de Montpellier II, 34095 Montpellier cedex 5,2 France
Received 25 June 2001/Returned for modification 18 July
2001/Accepted 13 August 2001
RhoG is a member of the Rho family of GTPases that activates
Rac1 and Cdc42 through a microtubule-dependent pathway. To gain understanding of RhoG downstream signaling, we performed a yeast two-hybrid screen from which we identified kinectin, a 156-kDa protein
that binds in vitro to conventional kinesin and enhances microtubule-dependent kinesin ATPase activity. We show that
RhoGGTP specifically interacts with the central domain of
kinectin, which also contains a RhoA binding domain in its C terminus.
Interaction was confirmed by coprecipitation of kinectin with active
RhoGG12V in COS-7 cells. RhoG, kinectin, and kinesin
colocalize in REF-52 and COS-7 cells, mainly in the endoplasmic
reticulum but also in lysosomes. Kinectin distribution in REF-52 cells
is modulated according to endogenous RhoG activity. In addition, by
using injection of anti-kinectin antibodies that challenge
RhoG-kinectin interaction or by blocking anti-kinesin antibodies, we
show that RhoG morphogenic activity relies on kinectin interaction and
kinesin activity. Finally, kinectin overexpression elicits Rac1- and
Cdc42-dependent cytoskeletal effects and switches cells to a RhoA
phenotype when RhoG activity is inhibited or microtubules are
disrupted. The functional links among RhoG, kinectin, and kinesin are
further supported by time-lapse videomicroscopy of COS-7 cells, which showed that the microtubule-dependent lysosomal transport is
facilitated by RhoG activation or kinectin overexpression and is
severely stemmed upon RhoG inhibition. These data establish that
kinectin is a key mediator of microtubule-dependent RhoG activity and
suggest that kinectin also mediates RhoG- and RhoA-dependent
antagonistic pathways.
*
Corresponding author. Mailing address: Centre de
Recherche en Biochimie Macromoléculaire, CNRS-UPR1086, 1919 Route
de Mende 34293, Montpellier cedex 5, France. Phone: 33 467613356. Fax: 33 467521559. E-mail: fort{at}crbm.cnrs-mop.fr.
Molecular and Cellular Biology, December 2001, p. 8022-8034, Vol. 21, No. 23
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.23.8022-8034.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Chandra Roy, B., Kakinuma, N., Kiyama, R.
(2009). Kank attenuates actin remodeling by preventing interaction between IRSp53 and Rac1. JCB
184: 253-267
[Abstract]
[Full Text]
-
Gupta, A., Decaillot, F. M., Gomes, I., Tkalych, O., Heimann, A. S., Ferro, E. S., Devi, L. A.
(2007). Conformation State-sensitive Antibodies to G-protein-coupled Receptors. J. Biol. Chem.
282: 5116-5124
[Abstract]
[Full Text]
-
Davies, S. L., Gibbons, C. E., Vizard, T., Ward, D. T.
(2006). Ca2+-sensing receptor induces Rho kinase-mediated actin stress fiber assembly and altered cell morphology, but not in response to aromatic amino acids. Am. J. Physiol. Cell Physiol.
290: C1543-C1551
[Abstract]
[Full Text]
-
Condliffe, A. M., Webb, L. M. C., Ferguson, G. J., Davidson, K., Turner, M., Vigorito, E., Manifava, M., Chilvers, E. R., Stephens, L. R., Hawkins, P. T.
(2006). RhoG Regulates the Neutrophil NADPH Oxidase. J. Immunol.
176: 5314-5320
[Abstract]
[Full Text]
-
Zhao, B., Maruo, S., Cooper, A., R. Chase, M., Johannsen, E., Kieff, E., Cahir-McFarland, E.
(2006). RNAs induced by Epstein-Barr virus nuclear antigen 2 in lymphoblastoid cell lines. Proc. Natl. Acad. Sci. USA
103: 1900-1905
[Abstract]
[Full Text]
-
Rabiner, C. A., Mains, R. E., Eipper, B. A.
(2005). Kalirin: A Dual Rho Guanine Nucleotide Exchange Factor That Is So Much More Than the Sum of Its Many Parts. Neuroscientist
11: 148-160
[Abstract]
-
Dransart, E., Morin, A., Cherfils, J., Olofsson, B.
(2005). Uncoupling of Inhibitory and Shuttling Functions of Rho GDP Dissociation Inhibitors. J. Biol. Chem.
280: 4674-4683
[Abstract]
[Full Text]
-
Govek, E.-E., Newey, S. E., Van Aelst, L.
(2005). The role of the Rho GTPases in neuronal development. Genes Dev.
19: 1-49
[Abstract]
[Full Text]
-
Blumenstein, L., Ahmadian, M. R.
(2004). Models of the Cooperative Mechanism for Rho Effector Recognition: IMPLICATIONS FOR RhoA-MEDIATED EFFECTOR ACTIVATION. J. Biol. Chem.
279: 53419-53426
[Abstract]
[Full Text]
-
de Toledo, M., Senic-Matuglia, F., Salamero, J., Uze, G., Comunale, F., Fort, P., Blangy, A.
(2003). The GTP/GDP Cycling of Rho GTPase TCL Is an Essential Regulator of the Early Endocytic Pathway. Mol. Biol. Cell
14: 4846-4856
[Abstract]
[Full Text]
-
Ma, X.-M., Huang, J., Wang, Y., Eipper, B. A., Mains, R. E.
(2003). Kalirin, a Multifunctional Rho Guanine Nucleotide Exchange Factor, Is Necessary for Maintenance of Hippocampal Pyramidal Neuron Dendrites and Dendritic Spines. J. Neurosci.
23: 10593-10603
[Abstract]
[Full Text]
-
Prieto-Sanchez, R. M., Bustelo, X. R.
(2003). Structural Basis for the Signaling Specificity of RhoG and Rac1 GTPases. J. Biol. Chem.
278: 37916-37925
[Abstract]
[Full Text]
-
Ong, L.-L., Er, C. P. N., Ho, A., Aung, M. T., Yu, H.
(2003). Kinectin Anchors the Translation Elongation Factor-1{delta} to the Endoplasmic Reticulum. J. Biol. Chem.
278: 32115-32123
[Abstract]
[Full Text]
-
Wennerberg, K., Ellerbroek, S. M., Liu, R.-Y., Karnoub, A. E., Burridge, K., Der, C. J.
(2002). RhoG Signals in Parallel with Rac1 and Cdc42. J. Biol. Chem.
277: 47810-47817
[Abstract]
[Full Text]
-
Charrasse, S., Meriane, M., Comunale, F., Blangy, A., Gauthier-Rouviere, C.
(2002). N-cadherin-dependent cell-cell contact regulates Rho GTPases and {beta}-catenin localization in mouse C2C12 myoblasts. JCB
158: 953-965
[Abstract]
[Full Text]
-
Tran, H., Pankov, R., Tran, S. D., Hampton, B., Burgess, W. H., Yamada, K. M.
(2002). Integrin clustering induces kinectin accumulation. J. Cell Sci.
115: 2031-2040
[Abstract]
[Full Text]