Phospho-regulation of the K-Cl cotransporter KCC2
Final Report Abstract
In this proposal we mainly analysed the impact of several bona fide phosphorylation sites on the regulation of KCC2 activity (WP3). We identified two novel regulatory phosphorylation sites (Ser932, Thr1008) that regulates the activity of KCC2. Furthermore, we detected phospho-sites which were directly targeted by staurosporine and NEM (Thr906, Ser932, Ser940, Thr1007 and Thr1008). We also showed that some phosphorylation sites (Ser31, Thr34, Thr934, Ser937, Thr999, Ser932) are involved in staurosporine and NEM mediated action and identified the underlying signalling pathway and kinases. Altogether, these analyses demonstrate that phospho-regulation of KCC2 activity is highly complex. In addition, we were able to generate Thr934Ala/Ser937Ala and KCC2 Thr934Ala/Ser937Asp transgenic mouse lines using the CRISPR/Cas system. Analysis of these mice point to a regulatory role of Ser937 in vivo. Furthermore, these mouse lines will provide excellent tools to gain further insight into the physiological role of KCC2-specific phospho-sites.
Publications
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(2018) Phosphoregulation of the intracellular termini of K-Cl cotransporter 2 (KCC2) enables flexible control of its activity. Journal of Biological Chemistry 293(44) 16984–16993
Cordshagen, A., Busch, W., Winkelhofer, M., Nothwang, H. G., Hartmann, A.-M.
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(2020) Staurosporine and NEM mainly impair WNK/SPAK/OSR1 mediated phosphorylation of KCC2 and NKCC1. PlosOne 15(5):e0232967
Zhang, J, Cordshagen, A., Medina, I., Nothwang, H. G., Wisniewski, J. R., Winklhofer, M., Hartmann, A.-M.
