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Akt3 is a privileged first responder in isozyme-specific electrophile response

Nature Chemical Biology volume 13, pages 333–338 (2017)Cite this article

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Abstract

Isozyme-specific post-translational regulation fine tunes signaling events. However, redundancy in sequence or activity renders links between isozyme-specific modifications and downstream functions uncertain. Methods to study this phenomenon are underdeveloped. Here we use a redox-targeting screen to reveal that Akt3 is a first-responding isozyme sensing native electrophilic lipids. Electrophile modification of Akt3 modulated downstream pathway responses in cells and Danio rerio (zebrafish) and markedly differed from Akt2-specific oxidative regulation. Digest MS sequencing identified Akt3 C119 as the privileged cysteine that senses 4-hydroxynonenal. A C119S Akt3 mutant was hypomorphic for all downstream phenotypes shown by wild-type Akt3. This study documents isozyme-specific and chemical redox signal–personalized physiological responses.

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Figure 1: Akt3 is a first-responding isozyme to reactive native lipid signals.
Figure 2: C119 of Akt3 is the unique HNE-sensing residue.
Figure 3: Lipid signals downregulate Akt3 kinase activity selectively.
Figure 4: Akt3(C119)-specific HNEylation upregulates transcriptional activities of tumor suppressor genes.

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Acknowledgements

We thank J. Fetcho and B. Miller (Cornell University) for scientific and technical guidance on zebrafish studies; J. Zhang (University of California, San Diego) and Johns Hopkins University for providing AktAR reporter plasmid; J. Poganik for his initial contributions to zebrafish studies; the zebrafish husbandry and microinjection/imaging facility (NIH R01 NS026593, J. Fetcho (Cornell University)), the Cornell NMR facility (NSF MRI: CHE-1531632, Y.A.), the Cornell Imaging Center (NIH 1S10RR025502, R.M. Williams (Cornell University)) and the Cornell proteomics and MS facility (NIH 1S10RR025449-01, S.Z.) for instrumentation. Supported by NIH New Innovator (1DP2GM114850), Beckman Young Investigator, NSF CAREER (CHE-1351400), Burroughs Wellcome CRTG and Sloan Fellowship programs (to Y.A.); HHMI predoctoral fellowship (59108350 to S.P.) and a Hill undergraduate fellowship (to S.L.S.) for student support.

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Author notes

  1. Marcus J C Long and Saba Parvez: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry and Chemical Biology, Cornell University, Ithaca, New York, USA

    Marcus J C Long, Saba Parvez, Yi Zhao, Sanjna L Surya, Yiran Wang & Yimon Aye

  2. Proteomics and Mass Spectrometry Facility, Institute of Biotechnology, Cornell University, Ithaca, New York, USA

    Sheng Zhang

  3. Department of Biochemistry, Weill Cornell Medicine, New York, New York, USA

    Yimon Aye

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  1. Marcus J C Long
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Contributions

M.J.C.L., S.P. and Y.A. designed the experiments. M.J.C.L. performed in vivo experiments in fish embryos. S.P. and M.J.C.L. performed cell-based experiments and collected the biochemical data from fish samples. Y.Z. and Y.W. performed chemical synthesis. S.L.S. assisted M.J.C.L. and S.P. with gene cloning and protein purification. S.Z. assisted in LC-MS/MS data analysis. M.J.C.L., S.P. and Y.A. analyzed and interpreted the data. Y.A. wrote the paper with editing contributions from M.J.C.L. and S.P.

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Correspondence to Yimon Aye.

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The authors declare no competing financial interests.

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Supplementary Results, Supplementary Tables 1–4 and Supplementary Figures 1–18. (PDF 4926 kb)

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Long, M., Parvez, S., Zhao, Y. et al. Akt3 is a privileged first responder in isozyme-specific electrophile response. Nat Chem Biol 13, 333–338 (2017). https://doi.org/10.1038/nchembio.2284

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