Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling

Y. Lu, M. Muller, D. Smith, B. Dutta, K. Komurov, S. Iadevaia, D. Ruths, J. T. Tseng, S. Yu, Q. Yu, L. Nakhleh, G. Balazsi, J. Donnelly, M. Schurdak, S. Morgan-Lappe, S. Fesik, P. T. Ram, G. B. Mills

    Research output: Contribution to journalArticle

    52 Scopus citations

    Abstract

    To identify regulators of intracellular signaling, we targeted 541 kinases and kinase-related molecules with small interfering RNAs (siRNAs), and determined their effects on signaling with a functional proteomics reverse-phase protein array (RPPA) platform assessing 42 phospho and total proteins. The kinome-wide screen demonstrated a strong inverse correlation between phosphorylation of AKT and mitogen-activated protein kinase (MAPK) with 115 genes that, when targeted by siRNAs, demonstrated opposite effects on MAPK and AKT phosphorylation. Network-based analysis identified the MAPK subnetwork of genes along with p70S6K and FRAP1 as the most prominent targets that increased phosphorylation of AKT, a key regulator of cell survival. The regulatory loops induced by the MAPK pathway are dependent on tuberous sclerosis complex 2 but demonstrate a lesser dependence on p70S6K than the previously identified FRAP1 feedback loop. The siRNA screen also revealed novel bi-directionality in the AKT and GSK3 (Glycogen synthase kinase 3) interaction, whereby genetic ablation of GSK3 significantly blocks AKT phosphorylation, an unexpected observation as GSK3 has only been predicted to be downstream of AKT. This method uncovered novel modulators of AKT phosphorylation and facilitated the mapping of regulatory loops.

    Original languageEnglish (US)
    Pages (from-to)4567-4577
    Number of pages11
    JournalOncogene
    Volume30
    Issue number45
    DOIs
    StatePublished - Nov 10 2011

    Keywords

    • AKT
    • MAPK
    • proteomics
    • siRNA
    • signaling networks

    ASJC Scopus subject areas

    • Molecular Biology
    • Genetics
    • Cancer Research

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  • Cite this

    Lu, Y., Muller, M., Smith, D., Dutta, B., Komurov, K., Iadevaia, S., Ruths, D., Tseng, J. T., Yu, S., Yu, Q., Nakhleh, L., Balazsi, G., Donnelly, J., Schurdak, M., Morgan-Lappe, S., Fesik, S., Ram, P. T., & Mills, G. B. (2011). Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling. Oncogene, 30(45), 4567-4577. https://doi.org/10.1038/onc.2011.164