Spatio-temporal cell signaling dynamics: towards an Akt subcellular localization fingerprint

MATÍAS BLAUSTEIN; ALEJANDRO COLMAN-LERNER

Rosario

Congreso; L Reunión Anual Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2014

SAIB

Cell-to-cell variation (CCV), also known as phenotypic heterogeneity, is a commonly observed phenomenon in normal and pathological conditions. Cancer is perhaps the most challenging situation in which to investigate what information is contained in CCV and how to use this information in therapy. Deregulation of Akt has been widely linked to cancer. This kinase orchestrates many biological functions including cell proliferation and survival. Our leading hypothesis is that the information contained in the expression level, posttranslational modifications, activity and specific subcellular localization of each Akt isoform can explain and predict variation in Akt target specificity, cell fate choices as well as in anti-tumor drug sensitivity displayed by individual cells. To test this hypothesis, we have developed immunofluorescence technics and designed fluorescent reporters to study CCV in the spatio-temporal dynamics of the Akt pathway. Here we describe a protocol to automatedly image and quantitatively measure and analyze the pattern of CCV in what we call the Akt subcellular localization fingerprint. We found novel compartments were Akt can be recruited and are now analyzing the molecular mechanisms that govern relocalization to these compartments, which in turn regulate specific crosstalk with other signaling pathways.