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Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling
To identify regulators of intracellular signaling we targeted 541 kinases and kinase-related molecules with 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 demons...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3175328/ https://www.ncbi.nlm.nih.gov/pubmed/21666717 http://dx.doi.org/10.1038/onc.2011.164 |
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author | Lu, Yiling Muller, Melissa Smith, Debra Dutta, Bhaskar Komurov, Kakajan Iadevaia, Sergio Ruths, Derek Tseng, Jen-Te Yu, Shuangxing Yu, Qinghua Nakhleh, Luay Balazsi, Gabor Donnelly, Jennifer Schurdak, Mark Morgan-Lappe, Susan Fesik, Stephen Ram, Prahlad T. Mills, Gordon B. |
author_facet | Lu, Yiling Muller, Melissa Smith, Debra Dutta, Bhaskar Komurov, Kakajan Iadevaia, Sergio Ruths, Derek Tseng, Jen-Te Yu, Shuangxing Yu, Qinghua Nakhleh, Luay Balazsi, Gabor Donnelly, Jennifer Schurdak, Mark Morgan-Lappe, Susan Fesik, Stephen Ram, Prahlad T. Mills, Gordon B. |
author_sort | Lu, Yiling |
collection | PubMed |
description | To identify regulators of intracellular signaling we targeted 541 kinases and kinase-related molecules with 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 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 TSC2 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 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. |
format | Online Article Text |
id | pubmed-3175328 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
record_format | MEDLINE/PubMed |
spelling | pubmed-31753282012-05-10 Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling Lu, Yiling Muller, Melissa Smith, Debra Dutta, Bhaskar Komurov, Kakajan Iadevaia, Sergio Ruths, Derek Tseng, Jen-Te Yu, Shuangxing Yu, Qinghua Nakhleh, Luay Balazsi, Gabor Donnelly, Jennifer Schurdak, Mark Morgan-Lappe, Susan Fesik, Stephen Ram, Prahlad T. Mills, Gordon B. Oncogene Article To identify regulators of intracellular signaling we targeted 541 kinases and kinase-related molecules with 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 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 TSC2 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 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. 2011-06-13 2011-11-10 /pmc/articles/PMC3175328/ /pubmed/21666717 http://dx.doi.org/10.1038/onc.2011.164 Text en Users may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms |
spellingShingle | Article Lu, Yiling Muller, Melissa Smith, Debra Dutta, Bhaskar Komurov, Kakajan Iadevaia, Sergio Ruths, Derek Tseng, Jen-Te Yu, Shuangxing Yu, Qinghua Nakhleh, Luay Balazsi, Gabor Donnelly, Jennifer Schurdak, Mark Morgan-Lappe, Susan Fesik, Stephen Ram, Prahlad T. Mills, Gordon B. Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling |
title | Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling |
title_full | Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling |
title_fullStr | Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling |
title_full_unstemmed | Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling |
title_short | Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling |
title_sort | kinome sirna-phosphoproteomic screen identifies networks regulating akt signaling |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3175328/ https://www.ncbi.nlm.nih.gov/pubmed/21666717 http://dx.doi.org/10.1038/onc.2011.164 |
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