Tumor suppressor p53 restricts Ras stimulation of RhoA and cancer cell motility

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Abstract

Many features of the cancer cell phenotype emerge as a result of cooperation between multiple oncogenic mutations. Here we show that activated RasV12 and loss of p53 function can cooperate to promote cell motility, a feature closely associated with cancer progression to malignancy. Our analysis indicates that RasV12 and loss of p53 synergistically induce RhoA activity, revealing a previously unknown role for p53 in tumor suppression. p53 prevents activation of RhoA and thus induction of cell motility by RasV12 through a simple signaling circuit, which integrates multiple inputs that converge on RhoA. Our data suggest that p53 suppresses cancer progression to malignancy by modulating the quality of Ras signaling.

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Figure 1: Activated Ras and mutant p53 cooperatively induce cell motility in mouse colon cells.
Figure 2: Cooperative activation of RhoA by Ras and mutant p53.
Figure 3: Oncogene-dependent RhoA activation contributes to increased cell motility in mp53/Ras cells.
Figure 4: Ras positively and negatively regulates RhoA activation by increasing RhoA membrane localization and stimulating p190 RhoGAP activity, respectively.
Figure 5: Negative regulation of RhoA by activated Ras mediated by p190 RhoGAP.
Figure 6: Loss of p53 function induces reduction of p190 RhoGAP tyrosine phosphorylation and accumulation of RhoA-GTP as well as motility in cancer cells expressing activated Ras.
Figure 7: Tumor suppressor p53 inhibits activation of RhoA and cell motility by RasV12 via 'AND' logic circuit.

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Acknowledgements

We thank L. Newman for expert assistance, D. Bohmann, H. McMurray, M. Noble, Y. Sun and J. Zhao for helpful discussions and L. Deleu for valuable help in the initial phase of the project. We also thank G. Evan (University of California, San Francisco) for providing anti–c-Myc (9E10), J. Collard (Netherlands Cancer Institute) for providing pGEX3X-GST-C21 and pGEX2TK-GST-PAKCD, A.J. Ridley (Ludwig Institute, London) for providing the RhoN19-Myc DNA fragment, J. Settleman (Massachusetts General Hospital Cancer Center) for providing HA-tagged p190 RhoGAP expression constructs, E. Sahai (Cancer Research UK) for providing pGEX-KG Tat-C3, R. Whitehead and A.W. Burgess (Ludwig Institute, Melbourne) for providing YAMC cells, B. Vogelstein (Johns Hopkins University) for providing p53-deficient HCT116 cells and G. Nolan (Stanford University) for providing Phoenix cells. This work was supported by the James P. Wilmot Foundation and by US National Institutes of Health grants CA90663 and GM075299.

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Correspondence to Hartmut Land.

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Supplementary information

Supplementary Fig. 1

p190 RhoGAP knockdown does not alter subcellular distribution of RhoA protein in the presence of RasV12 (PDF 562 kb)

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Xia, M., Land, H. Tumor suppressor p53 restricts Ras stimulation of RhoA and cancer cell motility. Nat Struct Mol Biol 14, 215–223 (2007) doi:10.1038/nsmb1208

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