Histone deacetylase 5 is a phosphorylation substrate of protein kinase D in osteoclasts.
Histone deacetylase 5 is a phosphorylation substrate of protein kinase D in osteoclasts.
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DOI：
10.1016/j.bone.2022.116393
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发表时间：
2022-06
期刊：
影响因子：
4.1
通讯作者：
Jensen, Eric D.
中科院分区：
文献类型：
--
作者：
Meyers, Carina Mello Guimaraes;Burciaga, Samuel D.;Faulkner, Bora;Kazemi, Parandis;Cohn, Jacob M.;Mansky, Kim C.;Jensen, Eric D.
关键词：
蛋白激酶D（PRKD）家族激酶是破骨细胞形成和功能所必需的。然而，破骨细胞中PRKD的底物是未知的。为了鉴定破骨细胞中的PRKD依赖性蛋白磷酸化，我们对用PRKD抑制剂CRT 0066101处理后破骨细胞中显示差异磷酸化的蛋白进行了定量LC-MS/MS磷酸蛋白质组学筛选。我们确定了757磷酸肽显示显着变化后PRKD抑制。在这些变化中，我们发现一组13种蛋白在PRKD共有磷酸化基序处显示磷酸化降低。该组包括组蛋白脱乙酰基酶5（HDAC 5），其是先前验证的PRKD靶标。考虑到这种已知的相互作用，工作表明HDAC可能是破骨细胞的重要调节因子，并且研究表明HDAC之间存在潜在的功能冗余，我们进一步研究了破骨细胞中PRKD和IIa类HDAC之间的关系。我们证实，CRT 0066101抑制内源性HDAC 5的磷酸化，并在较小程度上抑制HDAC 4的磷酸化，而HDAC 7磷酸化不受影响。从Hdac 5全基因敲除小鼠破骨细胞培养物显示受损的分化和降低的再吸收骨的能力，而破骨细胞中的Hdac 4的条件性敲除在体外或体内没有表现出表型。在Hdac 5 KO破骨细胞中，CRT 0066101的抑制作用降低。总之，这些数据表明，PRKD/HDAC 5轴有助于破骨细胞的形成在体外，并表明，这一途径可能有助于调节骨骼动力学在体内。
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.
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影响因子：
5.6
作者：
Leightner, Amanda C.;Meyers, Carina Mello Guimaraes;Jensen, Eric D.
通讯作者：
Jensen, Eric D.
影响因子：
4
作者：
Algate, Kent;Haynes, David;Cantley, Melissa
通讯作者：
Cantley, Melissa
影响因子：
7.3
作者：
Ho, Terence C. S.;Chan, Alex H. Y.;Ganesan, A.
通讯作者：
Ganesan, A.
影响因子：
4.8
作者：
Mansky, Kim C.;Jensen, Eric D.;Gopalakrishnan, Rajaram
通讯作者：
Gopalakrishnan, Rajaram
影响因子：
5.3
作者：
Chang, SR;McKinsey, TA;Olson, EN
通讯作者：
Olson, EN