PSKH1 kinase activity is differentially modulated via allosteric binding of Ca <sup>2+</sup> sensor proteins

C Christopher R. Horne (Walter and Eliza Hall Institute of Medical Research) T Toby A. Dite (Walter and Eliza Hall Institute of Medical Research) S Samuel N. Young (Walter and Eliza Hall Institute of Medical Research) L Lucy J. Mather (Walter and Eliza Hall Institute of Medical Research) L Laura F. Dagley (Walter and Eliza Hall Institute of Medical Research) J Jared L. Johnson (Meyer Cancer Center, Weill Cornell Medicine) T Tomer M. Yaron-Barir (Meyer Cancer Center, Weill Cornell Medicine) E Emily M. Huntsman (Meyer Cancer Center, Weill Cornell Medicine) L Leonard A. Daly (Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool) D Dominic P. Byrne (Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool) A Antonia L. Cadell (Cancer Ecosystems Program, Garvan Institute of Medical Research) B Boaz H. Ng (Cancer Ecosystems Program, Garvan Institute of Medical Research) J Jumana Yousef (Walter and Eliza Hall Institute of Medical Research) D Dylan H. Multari (Walter and Eliza Hall Institute of Medical Research) L Lianju Shen (Walter and Eliza Hall Institute of Medical Research) L Luke M. McAloon (Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University) G Gerard Manning (NuaBio Research) M Mark A. Febbraio A Anthony R. Means (Molecular and Cellular Biology, Baylor College of Medicine) L Lewis C. Cantley M Maria C. Tanzer D David R. Croucher (Cancer Ecosystems Program, Garvan Institute of Medical Research) C Claire E. Eyers (Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool) P Patrick A. Eyers (Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool) J John W. Scott (Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University) J James M. Murphy

Abstract

Protein Serine Kinase H1 (PSKH1) was recently identified as a crucial factor in kidney development and is overexpressed in prostate, lung, and kidney cancers. However, little is known about PSKH1 regulatory mechanisms, leading to its classification as a “dark” kinase. Here, we used biochemistry and mass spectrometry to define PSKH1’s consensus substrate motif, protein interactors, and how interactors, including Ca 2+ sensor proteins, promote or suppress activity. Intriguingly, despite the absence of a canonical Calmodulin binding motif, Ca 2+ -Calmodulin activated PSKH1 while, in contrast, the ER-resident Ca 2+ sensor of the Cab45, Reticulocalbin, Erc55, Calumenin (CREC) family, Reticulocalbin-3, suppressed PSKH1 catalytic activity. In addition to antagonistic regulation of the PSKH1 kinase domain by Ca 2+ sensing proteins, we identified UNC119B as a protein interactor that activates PSKH1 via direct engagement of the kinase domain. Our findings identify complementary allosteric mechanisms by which regulatory proteins tune PSKH1’s catalytic activity and raise the possibility that different Ca 2+ sensors may act more broadly to tune kinase activities by detecting and decoding extremes of intracellular Ca 2+ concentrations.

Article Details

Volume / Issue Vol. 122, Issue 8
Published February 25, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (26)

C

Christopher R. Horne

Walter and Eliza Hall Institute of Medical Research

T

Toby A. Dite

Walter and Eliza Hall Institute of Medical Research

S

Samuel N. Young

Walter and Eliza Hall Institute of Medical Research

L

Lucy J. Mather

Walter and Eliza Hall Institute of Medical Research

L

Laura F. Dagley

Walter and Eliza Hall Institute of Medical Research

J

Jared L. Johnson

Meyer Cancer Center, Weill Cornell Medicine

T

Tomer M. Yaron-Barir

Meyer Cancer Center, Weill Cornell Medicine

E

Emily M. Huntsman

Meyer Cancer Center, Weill Cornell Medicine

L

Leonard A. Daly

Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool

D

Dominic P. Byrne

Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool

A

Antonia L. Cadell

Cancer Ecosystems Program, Garvan Institute of Medical Research

B

Boaz H. Ng

Cancer Ecosystems Program, Garvan Institute of Medical Research

J

Jumana Yousef

Walter and Eliza Hall Institute of Medical Research

D

Dylan H. Multari

Walter and Eliza Hall Institute of Medical Research

L

Lianju Shen

Walter and Eliza Hall Institute of Medical Research

L

Luke M. McAloon

Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University

G

Gerard Manning

NuaBio Research

M

Mark A. Febbraio

A

Anthony R. Means

Molecular and Cellular Biology, Baylor College of Medicine

L

Lewis C. Cantley

M

Maria C. Tanzer

D

David R. Croucher

Cancer Ecosystems Program, Garvan Institute of Medical Research

C

Claire E. Eyers

Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool

P

Patrick A. Eyers

Department of Biochemistry, Cell and Systems Biology, Institute of Systems, Molecular and Integrative Biology, University of Liverpool

J

John W. Scott

Drug Discovery Biology, Monash Institute of Pharmaceutical Sciences, Monash University

J

James M. Murphy