Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling

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

doi:10.1038/onc.2011.164

Cited by 58 publications

(50 citation statements)

References 37 publications

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“…The mechanism by which depletion of GSK3β leads to the observed decrease in SRPK1 levels still remains to be determined, i.e., whether GSK3β acts directly on SRPK1 through phosphorylation or whether additional downstream effector proteins or protein complex formation are involved. For example, a genome-wide kinome screen revealed previously unpredicted networks regulated by GSK3β (Lu et al 2011).…”

Section: Discussionmentioning

confidence: 99%

Phosphorylation of SRSF1 by SRPK1 regulates alternative splicing of tumor-related Rac1b in colorectal cells

Gonçalves

Henriques

Pereira

et al. 2014

RNA

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The premessenger RNA of the majority of human genes can generate various transcripts through alternative splicing, and different tissues or disease states show specific patterns of splicing variants. These patterns depend on the relative concentrations of the splicing factors present in the cell nucleus, either as a consequence of their expression levels or of post-translational modifications, such as protein phosphorylation, which are determined by signal transduction pathways. Here, we analyzed the contribution of protein kinases to the regulation of alternative splicing variant Rac1b that is overexpressed in certain tumor types. In colorectal cells, we found that depletion of AKT2, AKT3, GSK3β, and SRPK1 significantly decreased endogenous Rac1b levels. Although knockdown of AKT2 and AKT3 affected only Rac1b protein levels suggesting a post-splicing effect, the depletion of GSK3β or SRPK1 decreased Rac1b alternative splicing, an effect mediated through changes in splicing factor SRSF1. In particular, the knockdown of SRPK1 or inhibition of its catalytic activity reduced phosphorylation and subsequent translocation of SRSF1 to the nucleus, limiting its availability to promote the inclusion of alternative exon 3b into the Rac1 pre-mRNA. Altogether, the data identify SRSF1 as a prime regulator of Rac1b expression in colorectal cells and provide further mechanistic insight into how the regulation of alternative splicing events by protein kinases can contribute to sustain tumor cell survival.

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Section: Discussionmentioning

confidence: 99%

Phosphorylation of SRSF1 by SRPK1 regulates alternative splicing of tumor-related Rac1b in colorectal cells

Gonçalves

Henriques

Pereira

et al. 2014

RNA

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“…However, the lack of predominant activating tyrosine kinase mutations in some epithelial cancers makes it difficult to pinpoint essential pathway drivers, and new approaches are needed to identify nonmutated, pathway-activated tyrosine kinases. To assess these kinases, phosphoproteomic enrichment strategies coupled with highly sensitive quantitative mass spectrometry (MS) and kinome-wide small interfering RNA (siRNA) screens have been utilized (37,(111)(112)(113)(114)(115)(116)(117)(118)(119)(120)(121)(122)(123)(124)(125).…”

Section: Phosphoproteomic and Kinome Sirna Screening Methods To Detecmentioning

confidence: 99%

“…These screens have identified kinases that behave as regulators of hormonal signaling (122), survival and proliferation (120), mechanisms of escape during hormonal depletion (126,127), regulators of downstream pathway activation (119,123), or mechanisms of drug resistance (118,121).…”

Section: Phosphoproteomic and Kinome Sirna Screening Methods To Detecmentioning

confidence: 99%

Clinical Targeting of Mutated and Wild-Type Protein Tyrosine Kinases in Cancer

Drake

Lee

Witte

2014

Molecular and Cellular Biology

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Clinical therapies for cancer have evolved from toxic, nontargeted agents to manageable, highly targeted therapies. Protein tyrosine kinases are a family of signaling molecules implicated in nearly every cancer type and are the foundation for the development of modern targeted agents. Recent genomic analyses have identified activating mutations, translocations, and amplifications of tyrosine kinases. Selective targeting of these genetically altered tyrosine kinases has resulted in significant clinical advances, including increased patient survival. This indicates that altered protein tyrosine kinases are the main drivers of many different cancers. However, lost during analyses of genetic lesions are the contributions of activated, wild-type kinases on tumordependent pathways. New approaches in phosphoproteomic technologies have identified several wild-type tyrosine kinase activation states, suggesting that non-genetically altered kinases can be essential "nodes" for signal transduction. Here, we summarize the evidence supporting the common mechanisms of protein tyrosine kinase activation in cancer and provide a personal perspective on the kinases BCR-ABL and BTK, as well as nonmutated kinase targets in prostate cancer, through our work. We outline the mechanisms of tyrosine kinase activation in the absence of direct mutation and discuss whether non-genetically altered tyrosine kinases or their associated downstream signaling pathways can be effectively targeted.T he seminal finding of BCR-ABL tyrosine kinase activity (1) and the subsequent development of the small-molecule inhibitor imatinib (2) have pioneered an era of therapies targeted toward mutated tyrosine kinases in cancer. Advancements in genomic sequencing and in our understanding of tumor signaling pathways have revealed several mutated and nonmutated pathway-activated tyrosine kinases. In this minireview, we will describe the different mechanisms of tyrosine kinase activation and how therapies tailored to these kinases have transformed disease outcomes. In particular, we will highlight two main examples of tyrosine kinases investigated in our lab, BCR-ABL and Bruton's tyrosine kinase (BTK). We will also examine a shifting paradigm in which nonmutated tyrosine kinases are being considered primary targets for tyrosine kinase inhibitor (TKI) therapy in cancer. Finally, we will discuss how phosphoproteomic approaches can identify activation of nonmutated kinase pathways and the challenges that we face by clinically targeting activated wild-type tyrosine kinases. TYROSINE KINASE SIGNALINGSince the discovery of v-SRC and v-ABL tyrosine phosphorylation 35 years ago (3, 4), considerable progress has been made in understanding how tyrosine phosphorylation contributes to normal cellular homeostasis and disease. Tyrosine phosphorylation is regulated by a family of enzymes known as tyrosine kinases. Tyrosine kinases are crucial mediators of normal cellular signal transduction functions, including cell proliferation, survival, migration, and apoptosis. Humans expr...

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“…6 In addition, a recent kinome-wide screen demonstrated that GSK-3 is a positive regulator of Akt, a key player in cancer cell survival and proliferation. 9 In addition to a direct role in cancer, GSK-3β can also indirectly promote cancer progression. Many human cancers arise and progress in the setting of chronic inflammatory states; thus, inflammatory conditions can initiate or promote oncogenic transformation.…”

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confidence: 99%

Glycogen synthase kinase (GSK)-3 and mammalian target of rapamycin complex 1 (mTORC1) cooperate to regulate protein S6 kinase 1 (S6K1)

2012

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Kinome siRNA-phosphoproteomic screen identifies networks regulating AKT signaling

Cited by 58 publications

References 37 publications

Phosphorylation of SRSF1 by SRPK1 regulates alternative splicing of tumor-related Rac1b in colorectal cells

Phosphorylation of SRSF1 by SRPK1 regulates alternative splicing of tumor-related Rac1b in colorectal cells

Clinical Targeting of Mutated and Wild-Type Protein Tyrosine Kinases in Cancer

Glycogen synthase kinase (GSK)-3 and mammalian target of rapamycin complex 1 (mTORC1) cooperate to regulate protein S6 kinase 1 (S6K1)

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