An endogenous PI3K interactome promoting astrocyte-mediated neuroprotection identifies a novel association with RNA-binding protein ZC3H14
| Autor: | Gerold Schmitt-Ulms, Samih Alqawlaq, Joseph D'Ercole, Anita H. Corbett, Declan Williams, Jeffrey L. Wrana, Alessandro Datti, Darren Chan, Sara W. Leung, Jeremy M Sivak, Alessandra Tuccitto, Izhar Livne-Bar |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
interactome Biochemistry Retinal ganglion Interactome Neuroprotection Poly(A)-Binding Proteins PI3K CFM cell-free media 03 medical and health sciences Phosphatidylinositol 3-Kinases astrocyte Growth factor receptor Tandem Mass Spectrometry medicine Immunoprecipitation ZC3H14 zinc finger CCCH-type containing 14 Molecular Biology THOC1 THO complex protein 1 PI3K/AKT/mTOR pathway ZC3h14 PDGFRA platelet-derived growth factor receptor A Phosphoinositide 3-kinase 030102 biochemistry & molecular biology biology PLA proximity ligation assay RNA-Binding Proteins poly(A) polyadenosine Cell Biology PDGF ACM astrocyte conditioned media RGC retinal ganglion cell Cell biology DNA-Binding Proteins 030104 developmental biology medicine.anatomical_structure PSM peptide-to-spectrum matches Astrocytes biology.protein neuroprotection RBP RNA-binding protein Chemical genetics PI3K phosphoinositide 3-kinase Astrocyte Research Article |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 1083-351X 0021-9258 |
| Popis: | Astrocytes can support neuronal survival through a range of secreted signals that protect against neurotoxicity, oxidative stress, and apoptotic cascades. Thus, analyzing the effects of the astrocyte secretome may provide valuable insight into these neuroprotective mechanisms. Previously, we characterized a potent neuroprotective activity mediated by retinal astrocyte conditioned media (ACM) on retinal and cortical neurons in metabolic stress models. However, the molecular mechanism underlying this complex activity in neuronal cells has remained unclear. Here, a chemical genetics screen of kinase inhibitors revealed phosphoinositide 3-kinase (PI3K) as a central player transducing ACM-mediated neuroprotection. To identify additional proteins contributing to the protective cascade, endogenous PI3K was immunoprecipitated from neuronal cells exposed to ACM or control media, followed by MS/MS proteomic analyses. These data pointed toward a relatively small number of proteins that coimmunoprecipitated with PI3K, and surprisingly only five were regulated by the ACM signal. These hits included expected PI3K interactors, such as the platelet-derived growth factor receptor A (PDGFRA), as well as novel RNA-binding protein interactors ZC3H14 (zinc finger CCCH-type containing 14) and THOC1 (THO complex protein 1). In particular, ZC3H14 has recently emerged as an important RNA-binding protein with multiple roles in posttranscriptional regulation. In validation studies, we show that PI3K recruitment of ZC3H14 is necessary for PDGF-induced neuroprotection and that this interaction is present in primary retinal ganglion cells. Thus, we identified a novel non-cell autonomous neuroprotective signaling cascade mediated through PI3K that requires recruitment of ZC3H14 and may present a promising strategy to promote astrocyte-secreted prosurvival signals. |
| Databáze: | OpenAIRE |
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