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Release of autoinhibition of ASEF by APC leads to CDC42 activation and tumor suppression

Nature Structural & Molecular Biology volume 14, pages 814–823 (2007)Cite this article

Abstract

Autoinhibition of the Rho guanine nucleotide exchange factor ASEF is relieved by interaction with the APC tumor suppressor. Here we show that binding of the armadillo repeats of APC to a 'core APC-binding' (CAB) motif within ASEF, or truncation of the SH3 domain of ASEF, relieves autoinhibition, allowing the specific activation of CDC42. Structural determination of autoinhibited ASEF reveals that the SH3 domain forms an extensive interface with the catalytic DH and PH domains to obstruct binding and activation of CDC42, and the CAB motif is positioned adjacent to the SH3 domain to facilitate activation by APC. In colorectal cancer cell lines, full-length, but not truncated, APC activates CDC42 in an ASEF-dependent manner to suppress anchorage-independent growth. We therefore propose a model in which ASEF acts as a tumor suppressor when activated by APC and inactivation of ASEF by mutation or APC truncation promotes tumorigenesis.

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Figure 1: ASEF is a GEF specific for CDC42.
Figure 2: ASEF induces filopodia in NIH3T3 fibroblasts.
Figure 3: ASEF is autoinhibited by the SH3 domain.
Figure 4: Identification of CAP motif in ASEF and ASEF2.
Figure 5: Differential activation of ASEF by full-length and truncated APC.
Figure 6: Structure of autoinhibited ASEF.
Figure 7: Detailed view of the SH3 domain interface.
Figure 8: ASEF is a candidate tumor suppressor.

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Acknowledgements

We thank D. Thiel and L. Bauer for technical assistance, Keith Burridge, (University of North Carolina) for the gift of GST-PAK RBD, the M. Hooker imaging facility at the University of North Carolina, Chapel Hill, and the staff at the SER-CAT beamlines for help with data collection. Use of the Advanced Photon Source was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences, under contract no. W-31-109-Eng-38. J.S. is supported by grants GM65533 and GM62299 from the US National Institutes of Health. C.J.D. is supported by grant CA063071 from the US National Institutes of Health. K.L.R. is supported by postdoctoral fellowship grant PF-05-129-01(GMC) from the American Cancer Society.

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Authors and Affiliations

  1. Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, Chapel Hill, 27599, North Carolina, USA

    Natalia Mitin, Channing J Der, John Sondek & Kent L Rossman

  2. Department of Pharmacology, University of North Carolina School of Medicine, Chapel Hill, 27599, North Carolina, USA

    Natalia Mitin, Laurie Betts, Marielle E Yohe, Channing J Der, John Sondek & Kent L Rossman

  3. Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, 27599, North Carolina, USA

    John Sondek

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  1. Natalia Mitin
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Contributions

N.M. and K.L.R. conceived, analyzed and performed experiments and wrote the manuscript. L.B. assisted with the structural determination of ASEF. M.E.Y. contributed ideas and assisted with construct design and protein purification. J.S. and C.J.D. contributed ideas and evaluated and discussed data.

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Correspondence to Channing J Der or Kent L Rossman.

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Mitin, N., Betts, L., Yohe, M. et al. Release of autoinhibition of ASEF by APC leads to CDC42 activation and tumor suppression. Nat Struct Mol Biol 14, 814–823 (2007). https://doi.org/10.1038/nsmb1290

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