Genetically encoded biosensors allow the noninvasive imaging of specific biochemical or biorecognition processes with the preservation of subcellular spatial and temporal information. Fused fluorescent proteins have revolutionized the ability of researchers to study protein localization and dynamics in live cells. Recently HMGB1 was discovered as one of the key regulators on the crosstalk of autophagy and apoptosis and the role of the protein in both processes is connected with its translocation from the nucleus to the cytoplasm. To investigate the role of HMGB1 in autophagic flux we engineered human A549 – lung adenocarcinoma (expressing wild type p53) and H1299 – non-small cell lung carcinoma (p53 negative) cells to express HMGB1 fused at the C-terminus of the green fluorescent protein (GFP). Coupled to fluorescence microscope for automated image acquisition of living cells this HMGB1-GFP-based biosensor is appropriate tool for monitoring the dynamic of HMGB1 translocation from the nucleus to the cytoplasm which is considered a hallmark for autophagy.
Key words: GFP-HMGB1 fused protein, lung cancer cells, autophagy