Open AccessMetastasis‐inducing protein S100A4 interacts with p53 in the nuclei of living cells
Author(s) -
Liu Zhen,
Meng Ying,
Wang Huijun,
Rudland Philip S.,
Barraclough Roger,
Zhang Shu
Publication year - 2019
Publication title -
precision radiation oncology
Language(s) - English
Resource type - Journals
ISSN - 2398-7324
DOI - 10.1002/pro6.63
Subject(s) - förster resonance energy transfer , cytoplasm , hela , microbiology and biotechnology , in vitro , bimolecular fluorescence complementation , green fluorescent protein , fluorescence , fluorescence recovery after photobleaching , photobleaching , biophysics , live cell imaging , hek 293 cells , fluorescence lifetime imaging microscopy , confocal microscopy , cell , yellow fluorescent protein , fluorescence microscope , chemistry , in vivo , biology , biochemistry , receptor , gene , membrane , physics , quantum mechanics
Objective S100A4 can bind to p53 in vitro . However, it is unclear whether the proteins interact in the nuclei of living cells. Methods Initially, using an optical biosensor, the ability of CFP‐S100A4 to bind with p53‐YFP was examined in vitro . The fluorescence resonance energy transfer (FRET) technique using laser confocal microscopy was applied to detect the interaction of S100A4 with p53 in living cells. Results Imaging FRET in HeLa cells expressing S100A4 and p53 showed an increase in cyan fluorescent protein fluorescence intensity, after photobleaching yellow fluorescent protein in cell nuclei. In comparison, the imaging of cell nuclei expressing mutant S100A4‐C and p53 or S100A4 and yellow fluorescent protein as the negative control showed the occurrence of FRET, and demonstrated that the interaction between S100A4 and p53 occurred in the nuclei of living HeLa cells. However, FRET did not occur in the cytoplasm of cells, showing that S100A4 did not interact with p53 in the cytoplasm. Conclusion S100A4 can interact directly with p53 in the nuclei of living cells. This might be a molecular basis for metastasis‐inducing S100A4 in vivo . The results also suggest that the inhibition of the calcium‐binding site of S100A4 might be a possible anticancer therapy.