WNK kinases sense molecular crowding and rescue cell volume via phase separation

Cary R Boyd-Shiwarski; Daniel J Shiwarski; Shawn E Griffiths; Rebecca T Beacham; Logan Norrell; Daryl E Morrison; Jun Wang; Jacob Mann; William Tennant; Eric N Anderson; Jonathan Franks; Michael Calderon; Kelly A Connolly; Muhammad Umar Cheema; Claire J Weaver; Lubika J Nkashama; Claire C Weckerly; Katherine E Querry; Udai Bhan Pandey; Christopher J Donnelly; Dandan Sun; Aylin R Rodan; Arohan R Subramanya

doi:10.1016/j.cell.2022.09.042

WNK kinases sense molecular crowding and rescue cell volume via phase separation

Cell. 2022 Nov 23;185(24):4488-4506.e20. doi: 10.1016/j.cell.2022.09.042. Epub 2022 Oct 31.

Authors

Cary R Boyd-Shiwarski¹, Daniel J Shiwarski², Shawn E Griffiths³, Rebecca T Beacham³, Logan Norrell⁴, Daryl E Morrison⁴, Jun Wang⁵, Jacob Mann⁶, William Tennant⁶, Eric N Anderson⁷, Jonathan Franks⁸, Michael Calderon⁸, Kelly A Connolly³, Muhammad Umar Cheema³, Claire J Weaver³, Lubika J Nkashama³, Claire C Weckerly⁹, Katherine E Querry³, Udai Bhan Pandey¹⁰, Christopher J Donnelly¹¹, Dandan Sun¹², Aylin R Rodan¹³, Arohan R Subramanya¹⁴

Affiliations

¹ Department of Medicine, Renal-Electrolyte Division, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA; Pittsburgh Center for Kidney Research, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
² Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA 15213, USA.
³ Department of Medicine, Renal-Electrolyte Division, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
⁴ Molecular Medicine Program, University of Utah, Salt Lake City, UT 84132, USA.
⁵ Department of Neurology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
⁶ Department of Neurobiology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
⁷ Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
⁸ Center for Biological Imaging, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
⁹ Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
¹⁰ Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA; Center for Protein Conformational Diseases, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
¹¹ Department of Neurobiology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA; Center for Protein Conformational Diseases, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
¹² Department of Neurology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA; VA Pittsburgh Healthcare System, Pittsburgh, PA 15240, USA.
¹³ Department of Human Genetics, University of Utah, Salt Lake City, UT 84132, USA; Molecular Medicine Program, University of Utah, Salt Lake City, UT 84132, USA; Department of Internal Medicine, Division of Nephrology and Hypertension, University of Utah, Salt Lake City, UT 84132, USA; Medical Service, VA Salt Lake City Health Care System, Salt Lake City, UT 84148, USA.
¹⁴ Department of Medicine, Renal-Electrolyte Division, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA; Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA; Center for Protein Conformational Diseases, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA; Pittsburgh Center for Kidney Research, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA; VA Pittsburgh Healthcare System, Pittsburgh, PA 15240, USA. Electronic address: [email protected].

Abstract

When challenged by hypertonicity, dehydrated cells must recover their volume to survive. This process requires the phosphorylation-dependent regulation of SLC12 cation chloride transporters by WNK kinases, but how these kinases are activated by cell shrinkage remains unknown. Within seconds of cell exposure to hypertonicity, WNK1 concentrates into membraneless condensates, initiating a phosphorylation-dependent signal that drives net ion influx via the SLC12 cotransporters to restore cell volume. WNK1 condensate formation is driven by its intrinsically disordered C terminus, whose evolutionarily conserved signatures are necessary for efficient phase separation and volume recovery. This disorder-encoded phase behavior occurs within physiological constraints and is activated in vivo by molecular crowding rather than changes in cell size. This allows kinase activity despite an inhibitory ionic milieu and permits cell volume recovery through condensate-mediated signal amplification. Thus, WNK kinases are physiological crowding sensors that phase separate to coordinate a cell volume rescue response.

Keywords: K-Cl cotransport; Na-K-2Cl cotransport; SLC12 cotransporter; WNK kinase; biomolecular condensates; cell volume regulation; hyperosmotic stress; macromolecular crowding; phase separation.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, Non-U.S. Gov't

MeSH terms

Cell Size
Phosphorylation
Protein Serine-Threonine Kinases*

Substances

Protein Serine-Threonine Kinases

Abstract

Publication types

MeSH terms

Substances

Grants and funding