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, Gordon Mills

Research output: Contribution to journalArticle

49 Citations (Scopus)

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
Externally publishedYes

Fingerprint

Small Interfering RNA
Mitogen-Activated Protein Kinases
Glycogen Synthase Kinase 3
Phosphorylation
70-kDa Ribosomal Protein S6 Kinases
Phosphotransferases
Protein Array Analysis
Tuberous Sclerosis
Proteomics
Genes
Cell Survival
Observation
Proteins

Keywords

  • AKT
  • MAPK
  • proteomics
  • signaling networks
  • siRNA

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

Cite this

Lu, Y., Muller, M., Smith, D., Dutta, B., Komurov, K., Iadevaia, S., ... Mills, G. (2011). Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling. Oncogene, 30(45), 4567-4577. https://doi.org/10.1038/onc.2011.164

Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling. / 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, Gordon.

In: Oncogene, Vol. 30, No. 45, 10.11.2011, p. 4567-4577.

Research output: Contribution to journalArticle

Lu, Y, Muller, M, Smith, D, Dutta, B, Komurov, K, Iadevaia, S, Ruths, D, Tseng, JT, Yu, S, Yu, Q, Nakhleh, L, Balazsi, G, Donnelly, J, Schurdak, M, Morgan-Lappe, S, Fesik, S, Ram, PT & Mills, G 2011, 'Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling', Oncogene, vol. 30, no. 45, pp. 4567-4577. https://doi.org/10.1038/onc.2011.164
Lu Y, Muller M, Smith D, Dutta B, Komurov K, Iadevaia S et al. Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling. Oncogene. 2011 Nov 10;30(45):4567-4577. https://doi.org/10.1038/onc.2011.164
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, Gordon. / Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling. In: Oncogene. 2011 ; Vol. 30, No. 45. pp. 4567-4577.
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