Analysis of cellular localization of РН domain of Bcr-Abl with USP1 protein and development of software for estimation of their phosphorylation sites

Abstract

Aim. This work was dedicated to investigation of cellular localization РН domain of Bcr-Abl with USP1 protein and to develop useful tool to aggregate information about protein phosphorylation sites. Methods. Mammalian cell transfection. Primers design. Widefield and STED fluorescent microscopy. Basic software development in Python. Results. Co-localization between USP1 and PH domain of Bcr-Abl has been detected. Primers for main domains of USP1 have been designed. Integration tool phospho-aggregate has been developed. Super-resolution image of intracellular distribution of PH domain of Bcr was obtained. Conclusions. Nuclear localization of USP1 in 293T cells and also point co-localizations of USP1 and РН domain of Bcr-Abl oncoloprotein show strong association of investigated proteins with specific cellular structures. Identification of these cellular structures can help to find out biological functions of of USP1 in CML. Tool phospho-aggregate can be used to simplify analysis and comparison of protein phosphorylation sites. Subdiffraction imaging of intracellular distribution of PH domain of Bcr shows that it clusters in structures that resemble shape of clathrin-coated vesicles.Keywords: Bcr-Abl, USP1, PH domain, cortactin, cellular localization, phosphorylation sites.