Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2011-11

AUTHORS

Yiling Lu, Melissa Muller, Debra Smith, Bhaskar Dutta, Kakajan Komurov, Sergio Iadevaia, Derek Ruths, Jen-Te Tseng, Shuangxing Yu, Qinghua Yu, Luay Nakhleh, Gabor Balazsi, Jennifer Donnelly, Mark Schurdak, Susan Morgan-Lappe, Stephen Fesik, Prahlad T. Ram, Gordon B. Mills

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. More... »

PAGES

4567

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/onc.2011.164

DOI

http://dx.doi.org/10.1038/onc.2011.164

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1011867058

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/21666717


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