Dual-specificity tyrosine-regulated kinases (DYRKs) auto-phosphorylate a critical tyrosine residue in their activation loop and phosphorylate their substrate on serine and threonine residues. The auto-phosphorylation occurs intramolecularly and is a one-off event. DYRK3 is selectively expressed at a high level in hematopoietic cells and attenuates erythroblast development, leading to anemia. In the present study, we determined the crystal structure of the mature form of human DYRK3 in complex with harmine, an ATP competitive inhibitor. The crystal structure revealed a phosphorylation site, residue S350, whose phosphorylation increases the stability of DYRK3 and enhances its kinase activity. In addition, our structural and biochemical assays suggest that the N-terminal auto-phosphorylation accessory domain stabilizes the DYRK3 protein, followed by auto-phosphorylation of the tyrosine of the activation loop, which is important for kinase activity. Finally, our docking analysis provides information for the design of novel and potent therapeutics to treat anemia.
Bibliographical noteFunding Information:
We thank the staff scientists of Pohang Light Source (South Korea) for assistance with the beamlines (5C and 7A) and of Photon Factory (Japan) beamlines (1A and 17A). We thank Soo Young Choi from Hallym University , who provided the PRAS40 gene. This work was supported by a National Research Foundation of Korea (NRF) grant funded by the Ministry of Science and ICT ( NRF-2016R1A2B2013305 , 2016R1A5A1010764 , 2017M3A9F6029755 ) and by the Strategic Initiative for Microbiomes in Agriculture and Food funded by Ministry of Agriculture, Food and Rural Affairs (Grant No. 916006-2 ).
All Science Journal Classification (ASJC) codes
- Structural Biology
- Molecular Biology