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A transforming mutation in the pleckstrin homology domain of AKT1 in cancer

Nature volume 448, pages 439–444 (2007)Cite this article

Abstract

Although AKT1 (v-akt murine thymoma viral oncogene homologue 1) kinase is a central member of possibly the most frequently activated proliferation and survival pathway in cancer, mutation of AKT1 has not been widely reported. Here we report the identification of a somatic mutation in human breast, colorectal and ovarian cancers that results in a glutamic acid to lysine substitution at amino acid 17 (E17K) in the lipid-binding pocket of AKT1. Lys 17 alters the electrostatic interactions of the pocket and forms new hydrogen bonds with a phosphoinositide ligand. This mutation activates AKT1 by means of pathological localization to the plasma membrane, stimulates downstream signalling, transforms cells and induces leukaemia in mice. This mechanism indicates a direct role of AKT1 in human cancer, and adds to the known genetic alterations that promote oncogenesis through the phosphatidylinositol-3-OH kinase/AKT pathway. Furthermore, the E17K substitution decreases the sensitivity to an allosteric kinase inhibitor, so this mutation may have important clinical utility for AKT drug development.

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Figure 1: Lys 17 alters the apo and Ins(1,3,4,5)P 4 -complexed AKT1 PHD structure.
Figure 2: The E17K mutation increases AKT1 activation in NIH 3T3 cells.
Figure 3: The E17K mutation alters AKT1 cellular localization.
Figure 4: Transformation of Rat1 fibroblasts by AKT1(E17K).
Figure 5: Timing and signature of AKT1(E17K)-driven leukaemia.

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Acknowledgements

We thank the Economic Development Board of Singapore for their support of this project through a RISC grant conferred to Lilly Research Laboratories. We also thank R. Gaynor for his comments, and W. Roeder for managing this project. We would like to thank the TGen DNA Sequencing Core, and are grateful to J. Tarrant for performing pathology examination of mouse blood smears, K. Neote and M. Swearingen for immunophenotyping, and P. Iversen for statistical analysis.

Author Contributions A.L.F. and C.H. contributed equally to this work. S.L.B. and R.S. crystallized the PHDs of AKT1. G.P.D. and D.J.Z performed the adoptive transfer studies. All authors discussed the results and commented on the manuscript.

X-ray crystallographic coordinates and structure factor files have been deposited in the Protein Data Bank (see Supplementary Table 2). E17K_APO has been assigned the code 2UZR for the coordinate entry. E17K_P4-inositiol has been assigned the code 2UZS for the coordinate entry.

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Authors and Affiliations

  1. Division of Integrated Cancer Genomics, Translational Genomics Research Institute, 445 N. Fifth Street, Phoenix, Arizona 85004, USA,

    John D. Carpten, Christiane M. Robbins, Galen Hostetter, Tracy Y. Moses, Stephanie Savage, Jeffrey Touchman & Michael Bittner

  2. Cancer Discovery Research,,

    Andrew L. Faber, Candice Horn, Gregory P. Donoho, Sophie Boguslawski, Mark Uhlik, Jian Du, Douglas J. Zeckner, Mei-Huei T. Lai, Kerry L. Blanchard & James E. Thomas

  3. Global Structural Biology,,

    Stephen L. Briggs & Richard Schevitz

  4. Integrative Biology, Lilly Research Laboratories, Eli Lilly & Company, Indianapolis, Indiana 46285, USA,

    Aimin Lin & Yue-Wei Qian

  5. Lilly Singapore Centre for Drug Discovery, 1 Science Park Road 04-01, The Capricorn, Singapore Science Park II, 117528 Singapore,

    Greg Tucker-Kellogg & Ketan Patel

  6. Pharmaceutical Genomics, Translational Genomics Research Institute, TGen Suite 110, 13208 E. Shea Boulevard, Scottsdale, Arizona 85259, USA,

    Spyro Mousses

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Correspondence to Kerry L. Blanchard or James E. Thomas.

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Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-6 with Legends, Supplementary Tables 1 -2, Legends for Supplementary Movies 1-5. (PDF 1307 kb)

Supplementary Video 1

This file contains Supplementary Video1 which shows the movement of GFP-PHwt in NIH3T3 cells over a period of several minutes after treatment with PDGF. (MOV 135 kb)

Supplementary Video 2

This file contains Supplementary Video 2 which shows the movement of GFP-PHE17K in NIH3T3 cells treated with PDGF over a period of several minutes. (MOV 234 kb)

Supplementary Video 3

This file contains Supplementary Video 3 which shows the movement of GFP-PHR25C in NIH3T3 cells over a period of several minutes after treatment with PDGF. (MOV 378 kb)

Supplementary Video 4

This file contains Supplementary Video 4 which shows the movement of GFP-PHwt in NIH3T3 cells over period of several minutes after treatment with LY294002 followed by treatment with PDGF. (MOV 387 kb)

Supplementary Video 5

This file contains Supplementary Video 5 which shows the movement of GFP-PHE17K in NIH3T3 cells over period of several minutes after treatment with LY294002 followed by treatment with PDGF. This legend was corrected on 18 July 2007. (MOV 286 kb)

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Carpten, J., Faber, A., Horn, C. et al. A transforming mutation in the pleckstrin homology domain of AKT1 in cancer. Nature 448, 439–444 (2007). https://doi.org/10.1038/nature05933

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