Histone deacetylase 5 is a phosphorylation substrate of protein kinase D in osteoclasts

Carina Mello Guimaraes Meyers, Samuel D. Burciaga, Bora Faulkner, Parandis Kazemi, Jacob M. Cohn, Kim C. Mansky, Eric D Jensen

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

Protein kinase D (PRKD) family kinases are required for formation and function of osteoclasts. However, the substrates of PRKD in osteoclasts are unknown. To identify PRKD-dependent protein phosphorylation in osteoclasts, we performed a quantitative LC-MS/MS phosphoproteomics screen for proteins showing differential phosphorylation in osteoclasts after treatment with the PRKD inhibitor CRT0066101. We identified 757 phosphopeptides showing significant changes following PRKD inhibition. Among the changes, we found a group of 13 proteins showing decreased phosphorylation at PRKD consensus phosphorylation motifs. This group includes histone deacetylase 5 (HDAC5), which is a previously validated PRKD target. Considering this known interaction, work suggesting HDACs may be important regulators of osteoclasts, and studies suggesting potential functional redundancy between HDACs, we further investigated the relationship between PRKD and class IIa HDACs in osteoclasts. We confirmed that CRT0066101 inhibits phosphorylation of endogenous HDAC5 and to a lesser extent HDAC4, whereas HDAC7 phosphorylation was not affected. Osteoclast cultures from Hdac5 global knockout mice displayed impaired differentiation and reduced ability to resorb bone, while conditional knockout of Hdac4 in osteoclasts showed no phenotype in vitro or in vivo. The inhibitory effect of CRT0066101 was reduced in Hdac5 KO osteoclasts. Together these data indicate that the PRKD/HDAC5 axis contributes to osteoclast formation in vitro and suggest that this pathway may contribute to regulation of skeletal dynamics in vivo.

Original languageEnglish (US)
Article number116393
JournalBone
Volume159
DOIs
StatePublished - Jun 2022

Bibliographical note

Funding Information:
This work was supported by a grant to EDJ and KM from the National Institute of Arthritis and Musculoskeletal and Skin Diseases ( NIH R01AR061352 ).

Publisher Copyright:
© 2022

Keywords

  • Bone remodeling
  • Histone deacetylases
  • Osteoclast differentiation
  • Protein kinase D
  • Protein phosphorylation

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