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PreprintCited by 8Microtubule and mitosis dynamicsGenomics and Chromatin DynamicsCancer-related Molecular Pathways

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Molecular requirements for PLK1 activation by T-loop phosphorylation

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Esposito Verza, Arianna, Conti, Duccio, Rodrigues Pedroso, Paulo D., Oberste-Lehn, Lina, Koerner, Carolin, Wohlgemuth, Sabine, Mansurkhodzhaev, Artem, Vetter, Ingrid R., Pesenti, Marion E., Musacchio, Andrea·2025·The EMBO Journal
DOI 10.1101/2025.07.25.666769PMC PMC12953774

Preprint — published as 10.1038/s44318-025-00681-0

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Abstract

Activation of PLK1, a master mitotic kinase, requires phosphorylation of its activation segment on Thr210, within a basic consensus sequence for Aurora kinases. Aurora B-dependent phosphorylation of Thr210 has been reported, but other evidence identified a strict requirement for the Aurora A partner Bora for Thr210 phosphorylation. Here, we investigate the elusive mechanistic basis for this requirement. We show that Aurora A:Bora phosphorylates Thr210 of PLK1 in vitro, while Aurora A, other Aurora A:activator complexes, and Aurora B:INCENP fail to target T210 even at high kinase/substrate ratios. A transient interaction of Bora and PLK1, identified by structural modelling and probed mutationally, is uniquely required for Thr210 phosphorylation. Dependency on Bora for Thr210 phosphorylation is eliminated after mutating Lys208, in the Aurora consensus, into arginine. This conservative mutation turns PLK1 into an excellent substrate of nearly all tested active Aurora kinases, especially Aurora B. Collectively, these results shine a new light on the specificity of the PLK1 activation mechanism.

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Metadata

Authors
Esposito Verza, A., Conti, D., Rodrigues Pedroso, P. D., Oberste-Lehn, L., Koerner, C., Wohlgemuth, S., Mansurkhodzhaev, A., Vetter, I. R., Pesenti, M. E., & Musacchio, A.
Affiliations
Max Planck Institute of Molecular Physiology (DE)
Corresponding author
Musacchio, Andrea
ORCID
Venue
The EMBO Journal
Publisher
openRxiv
Published
2025-07-29
Volume
45
Issue
5
Pages
1496-1536
ISSN
0261-4189 (ISSN-L)
DOI
10.1101/2025.07.25.666769
PMC
PMC12953774
OpenAlex
W7125977885
Cited by
8
References
1
Citations by year
2026: 7 · 2025: 1
FWCI
23.46
Type
journal article
Language
en
Open access
Licence
Locations
  • The EMBO Journal (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • Universitätsbibliographie, Universität Duisburg-Essen (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • MPG.PuRe (Max Planck Society) (submittedVersion)
Funders
  • European Molecular Biology Organization (ALFT439-2019)
  • Deutsche Forschungsgemeinschaft (CRC 1430)
  • HORIZON EUROPE European Research Council (SyG 951430)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
  • Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
Keywords
PLK1, T-loop phosphorylation, AURKA, Borås, Aurora B kinase, INCENP, kinase substrate specificity, protein-protein interaction, mitosis
MeSH terms
Polo-Like Kinase 1, Enzyme Activation, Humans, Phosphorylation, Proto-Oncogene Proteins, Protein Serine-Threonine Kinases, Cell Cycle Proteins, Aurora Kinase A, Aurora Kinase B

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