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PreprintCited by 4Protein Kinase Regulation and GTPase SignalingProtein Structure and DynamicsS100 Proteins and Annexins

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Unconventional binding of Calmodulin to CHK2 kinase inhibits catalytic activity

Open access
Horne, Christopher R., Wang, Tingting, Young, Samuel N., Dite, Toby A., Nyvall, Hunter G., Suresh, Sushant, Davies, Katherine A., Mather, Lucy J., Dagley, Laura F., Manning, Gerard, Means, Anthony R., Burke, John E., Petersen, Janni, Scott, John W., Murphy, James M.·2024·Biochemical Journal
DOI 10.1101/2024.10.08.617309PMC PMC12751073

Preprint — published as 10.1042/bcj20253431

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Abstract

Calmodulin (CaM) serves an essential role in eukaryotic cells as a Ca 2+ sensor. Ca 2+ binding leads to conformation changes in CaM that enable engagement of a repertoire of enzymes and the regulation of their catalytic activities. Classically, Ca 2+ -CaM binds to an inhibitory pseudosubstrate sequence C-terminal to the kinase domain in members of the Ca 2+ -CaM dependent protein kinase (CAMK) family, and relieves inhibition to promote catalytic activity. Here, we report an unexpected mechanism by which CaM can bind CHK2 kinase to inhibit its kinase activity. Using biochemical, biophysical, structural mass spectrometry, and cellular approaches, we identify a direct interaction of Ca 2+ -CaM with the CHK2 kinase domain that suppresses CHK2 catalytic activity in vitro and is crucial for cell proliferation in human cells following DNA damage. Our findings add direct suppression of kinase activity to the repertoire of CaM’s functions, complementing the paradigmatic mechanism of promoting kinase activity through autoinhibitory domain sequestration.

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Metadata

Authors
Horne, C. R., Wang, T., Young, S. N., Dite, T. A., Nyvall, H. G., Suresh, S., Davies, K. A., Mather, L. J., Dagley, L. F., Manning, G., Means, A. R., Burke, J. E., Petersen, J., Scott, J. W., & Murphy, J. M.
Affiliations
  • The University of Melbourne (AU)
  • Flinders University (AU)
  • Walter and Eliza Hall Institute of Medical Research (AU)
  • University of Victoria (CA)
  • Baylor College of Medicine (US)
  • University of British Columbia (CA)
  • Florey Institute of Neuroscience and Mental Health (AU)
Corresponding author
  • Horne, Christopher R.
  • Scott, John W.
  • Murphy, James M.
ORCID
Venue
Biochemical Journal
Publisher
openRxiv
Published
2024-10-08
Volume
482
Issue
23
Pages
1759-1777
ISSN
0264-6021 (ISSN-L)
DOI
10.1101/2024.10.08.617309
PMC
PMC12751073
OpenAlex
W4414534112
Cited by
4
References
1
Citations by year
2026: 2 · 2025: 2
FWCI
1.20
Type
journal article
Language
en
Open access
Licence
Locations
  • Biochemical Journal (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • Digital Commons-TMC (Texas Medical Center) (submittedVersion, cc-by)
  • Minerva Access (University of Melbourne) (submittedVersion, cc-by)
Funders
  • Cancer Research Society (1052949)
  • Michael Smith Health Research BC (2020-04241, 17868)
  • Natural Sciences and Engineering Research Council of Canada (rgpin-2020-04241, 17868, 2020-04241)
  • Medical Research Council (17868)
  • Australian Research Council (DP210102840)
  • National Health and Medical Research Council (2034044, 2001817, 9000719, 1172929, 2034104, 9000719, 2034044, 2001817, 1172929, 2034104)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › S100 Proteins and Annexins
Keywords
calmodulin, Chk2, kinase inhibition, calcium signaling, pseudosubstrate, autoinhibition, DNA damage response, cell proliferation, protein-protein interaction, kinase domain
MeSH terms
Protein Domains, Calcium, Calmodulin, DNA Damage, Humans, Protein Binding, Cell Proliferation, Checkpoint Kinase 2

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