Cited by 339Protein Kinase Regulation and GTPase SignalingCellular transport and secretionMechanisms of cancer metastasis

Specificity and mechanism of action of EHT 1864, a novel small molecule inhibitor of Rac family small GTPases

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Shutes, Adam, Onesto, Cercina, Picard, Virginie, Leblond, Bertrand, Schweighoffer, Fabien, Der, Channing J.·2007·Journal of Biological Chemistry

Abstract

There is now considerable experimental evidence that aberrant activation of Rho family small GTPases promotes the uncontrolled proliferation, invasion, and metastatic properties of human cancer cells. Therefore, there is considerable interest in the development of small molecule inhibitors of Rho GTPase function. However, to date, most efforts have focused on inhibitors that indirectly block Rho GTPase function, by targeting either enzymes involved in post-translational processing or downstream protein kinase effectors. We recently determined that the EHT 1864 small molecule can inhibit Rac function in vivo. In this study, we evaluated the biological and biochemical specificities and biochemical mechanism of action of EHT 1864. We determined that EHT 1864 specifically inhibited Rac1-dependent platelet-derived growth factor-induced lamellipodia formation. Furthermore, our biochemical analyses with recombinant Rac proteins found that EHT 1864 possesses high affinity binding to Rac1, as well as the related Rac1b, Rac2, and Rac3 isoforms, and this association promoted the loss of bound nucleotide, inhibiting both guanine nucleotide association and Tiam1 Rac guanine nucleotide exchange factor-stimulated exchange factor activity in vitro. EHT 1864 therefore places Rac in an inert and inactive state, preventing its engagement with downstream effectors. Finally, we evaluated the ability of EHT 1864 to block Rac-dependent growth transformation, and we determined that EHT 1864 potently blocked transformation caused by constitutively activated Rac1, as well as Rac-dependent transformation caused by Tiam1 or Ras. Taken together, our results suggest that EHT 1864 selectively inhibits Rac downstream signaling and transformation by a novel mechanism involving guanine nucleotide displacement.

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  1. Gao, Y., Dickerson, J. B., Guo, F., Zheng, J., and Zheng, Y. (2004) Proc. Natl. Acad. Sci. U. S. A. 101, 7618 -7623
  2. Manne, V., Yamazaki, S., and Kung, H. F. (1984) Proc. Natl. Acad. Sci.

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Authors
Shutes, A., Onesto, C., Picard, V., Leblond, B., Schweighoffer, F., & Der, C. J.
Affiliations
University of North Carolina at Chapel Hill (US)
Corresponding author
Der, Channing J.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2007-12
Volume
282
Issue
49
Pages
35666-35678
ISSN
0021-9258 (ISSN-L)
OpenAlex
W2124054733
Cited by
336
References
62
Citations by year
2026: 14 · 2025: 18 · 2024: 22 · 2023: 16 · 2022: 18 · 2021: 21 · 2020: 22 · 2019: 20 · 2018: 16 · 2017: 22 · 2016: 20 · 2015: 21 · 2014: 7 · 2013: 27 · 2012: 25
FWCI
3.98
Type
journal article
Language
en
Open access
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
  • UNC Libraries
Funders
  • Susan G. Komen for the Cure
  • National Institutes of Health (CA063071, CA67771)
  • National Cancer Institute (CA 92240, CA 063071, R01CA063071, CA 67771, R01CA092240, U19CA067771)
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 › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mechanisms of cancer metastasis
Keywords
Rac1, guanine nucleotide exchange factor, Tiam1, lamellipodia formation, platelet-derived growth factor, cell transformation, small molecule inhibitors, metastasis
MeSH terms
T-Lymphoma Invasion and Metastasis-inducing Protein 1, Animals, Cell Transformation, Neoplastic, Enzyme Activation, Humans, Neoplasm Invasiveness, Neoplasm Metastasis, Neoplasms, Protein Processing, Post-Translational, Pseudopodia, Pyrones, Quinolines, Signal Transduction, Guanine Nucleotide Exchange Factors, rac GTP-Binding Proteins, NIH 3T3 Cells, Cell Line, Tumor, Cell Proliferation, Mice

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