Producción CyT
Biocell - Akt posttranslational and subcellular localization profiling: towards an Akt fingerprint

Congreso

Fecha
2015
Editorial y Lugar de Edición
Biocell
Resumen Información suministrada por el agente en SIGEVA
Cell-to-cell variability (CCV) is observed in normal and pathological conditions. Cancer is perhaps the most challenging situation in which to investigate how to use information present in CCV for therapy. Deregulation of Akt, a kinase that orchestrates many biological functions including cell proliferation and survival, has been widely linked to cancer. The information contained in the expression level, posttranslational modifications, activity and specific subcellular localization of each Akt... Cell-to-cell variability (CCV) is observed in normal and pathological conditions. Cancer is perhaps the most challenging situation in which to investigate how to use information present in CCV for therapy. Deregulation of Akt, a kinase that orchestrates many biological functions including cell proliferation and survival, has been widely linked to cancer. The information contained in the expression level, posttranslational modifications, activity and specific subcellular localization of each Akt isoform in each cell might explain differential Akt target specificity, CCV in cell fate choices as well as in anti-tumor drug sensitivity. We have developed immunofluorescence-based technics and designed fluorescent reporters to study CCV in the spatio-temporal dynamics of the Akt pathway. Here, we describe a protocol for automated imaging and quantitative measurement and analysis of the localization and phosphorylation profiles of Akt and its substrates. The combined dataset obtained defines what we call the ?Akt fingerprint?. We found novel subcellular compartments were Akt is recruited and we are now analyzing the molecular mechanisms that govern relocalization to these places. Analysis of Akt substrates by phosphorylation and localization profiles as well as classification of cancer cell lines by Akt fingerprint can lead us to understand complex cell and tumor behaviors.
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Palabras Clave
cell-to-cell variationsubcellular localizationcell signalingAkt