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PreprintCited by 5Protein Kinase Regulation and GTPase SignalingHeat shock proteins researchProtein Structure and Dynamics

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Evolutionary and cellular analysis of the dark pseudokinase PSKH2

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Byrne, Dominic P, Shrestha, Safal, Daly, Leonard A, Marensi, Vanessa, Ramakrishnan, Krithika, Eyers, Claire E, Kannan, Natarajan, Eyers, Patrick A·2022·Biochemical Journal
DOI 10.1101/2022.09.10.507278PMC PMC9988210

Preprint — published as 10.1042/bcj20220474 · Paper page

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Abstract

Pseudokinases, so named because they lack one or more conserved canonical amino acids that define their catalytically-active relatives, have evolved a variety of biological functions in both prokaryotic and eukaryotic organisms. Human PSKH2 is closely related to the canonical kinase PSKH1, which maps to the CAMK family of protein kinases. Primates encode PSKH2 in the form of a pseudokinase, which is predicted to be catalytically inactive due to loss of the invariant catalytic Asp residue. Although the biological role(s) of vertebrate PSKH2’s remains unclear, we previously identified species-level adaptions in PSKH2 that have led to the appearance of kinase or pseudokinase variants in vertebrate genomes alongside a canonical PSKH1 paralog. In this paper we confirm that, as predicted, PSKH2 lacks detectable protein phosphotransferase activity, and exploit structural informatics, biochemistry and cellular proteomics to begin to characterise vertebrate PSKH2 orthologues. AlphaFold 2-based structural analysis predicts functional roles for both the PSKH2 N- and C-regions that flank the pseudokinase domain core, and cellular truncation analysis confirms that the N-terminal domain, which contains a conserved myristoylation site, is required for both stable human PSKH2 expression and localisation to a membrane-rich subcellular fraction containing mitochondrial proteins. Using mass spectrometry-based proteomics, we confirm that human PSKH2 is part of a cellular mitochondrial protein network, and that its expression is regulated through client-status within the HSP90/Cdc37 molecular chaperone system. HSP90 interactions are mediated through binding to the PSKH2 C-terminal tail, leading us to predict that this region might act as both a cis and trans regulatory element, driving outputs linked to the PSKH2 pseudokinase domain that are important for functional signalling.

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Metadata

Authors
Byrne, D. P., Shrestha, S., Daly, L. A., Marensi, V., Ramakrishnan, K., Eyers, C. E., Kannan, N., & Eyers, P. A.
Affiliations
  • University of Liverpool (GB)
  • University of Georgia (US)
Corresponding author
Kannan, Natarajan
ORCID
Venue
Biochemical Journal
Publisher
openRxiv
Published
2022-09-10
Volume
480
Issue
2
Pages
141-160
ISSN
0264-6021 (ISSN-L)
DOI
10.1101/2022.09.10.507278
PMC
PMC9988210
OpenAlex
W4311547358
Cited by
5
References
1
Citations by year
2026: 1 · 2025: 2 · 2024: 1 · 2023: 1
FWCI
0.33
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Biochemical Journal (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • North West Cancer Research (CR1208, CR1097)
  • National Institutes of Health (BB/N021703/1, BB/S018514/1)
  • Biotechnology and Biological Sciences Research Council (BB/N021703/1, BB/S018514/1, BB/N021703, BB/R000182/1)
  • National Cancer Institute (U01CA239106)
  • Common Fund (U01CA239106)
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 › Heat shock proteins research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
pseudokinase, protein kinase evolution, myristoylation, subcellular localization
MeSH terms
Animals, Biological Evolution, Humans, Phosphorylation, Protein Kinases, Signal Transduction, Molecular Chaperones, HSP90 Heat-Shock Proteins

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