TY - JOUR
T1 - Hydrostatic Pressure Sensing by WNK kinases
AU - Humphreys, John M.
AU - Teixeira, Liliana R.
AU - Akella, Radha
AU - He, Haixia
AU - Kannangara, Ashari R.
AU - Sekulski, Kamil
AU - Pleinis, John
AU - Liwocha, Joanna
AU - Jiou, Jenny
AU - Servage, Kelly A.
AU - Orth, Kim
AU - Joachimiak, Lukasz
AU - Rizo, Josep
AU - Cobb, Melanie H.
AU - Brautigam, Chad A.
AU - Rodan, Aylin R.
AU - Goldsmith, Elizabeth J.
N1 - Publisher Copyright:
© 2023 Humphreys et al.
PY - 2023/10/1
Y1 - 2023/10/1
N2 - Previous study has demonstrated that the WNK kinases 1 and 3 are direct osmosensors consistent with their established role in cell-volume control. WNK kinases may also be regulated by hydrostatic pressure. Hydrostatic pressure applied to cells in culture with N2 gas or to Drosophila Malpighian tubules by centrifugation induces phosphorylation of downstream effectors of endogenous WNKs. In vitro, the autophosphorylation and activity of the unphosphorylated kinase domain of WNK3 (uWNK3) is enhanced to a lesser extent than in cells by 190 kPa applied with N2 gas. Hydrostatic pressure measurably alters the structure of uWNK3. Data from size exclusion chromatography in line with multi-angle light scattering (SEC-MALS), SEC alone at different back pressures, analytical ultracentrifugation (AUC), NMR, and chemical crosslinking indicate a change in oligomeric structure in the presence of hydrostatic pressure from a WNK3 dimer to a monomer. The effects on the structure are related to those seen with osmolytes. Potential mechanisms of hydrostatic pressure activation of uWNK3 and the relationships of pressure activation to WNK osmosensing are discussed.
AB - Previous study has demonstrated that the WNK kinases 1 and 3 are direct osmosensors consistent with their established role in cell-volume control. WNK kinases may also be regulated by hydrostatic pressure. Hydrostatic pressure applied to cells in culture with N2 gas or to Drosophila Malpighian tubules by centrifugation induces phosphorylation of downstream effectors of endogenous WNKs. In vitro, the autophosphorylation and activity of the unphosphorylated kinase domain of WNK3 (uWNK3) is enhanced to a lesser extent than in cells by 190 kPa applied with N2 gas. Hydrostatic pressure measurably alters the structure of uWNK3. Data from size exclusion chromatography in line with multi-angle light scattering (SEC-MALS), SEC alone at different back pressures, analytical ultracentrifugation (AUC), NMR, and chemical crosslinking indicate a change in oligomeric structure in the presence of hydrostatic pressure from a WNK3 dimer to a monomer. The effects on the structure are related to those seen with osmolytes. Potential mechanisms of hydrostatic pressure activation of uWNK3 and the relationships of pressure activation to WNK osmosensing are discussed.
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U2 - 10.1091/MBC.E23-03-0090
DO - 10.1091/MBC.E23-03-0090
M3 - Article
C2 - 37585288
AN - SCOPUS:85171900476
SN - 1059-1524
VL - 34
JO - Molecular biology of the cell
JF - Molecular biology of the cell
IS - 11
M1 - ar109
ER -