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ID 57503
フルテキストURL
著者
Takabatake, Shota Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
Ohtsuka, Satomi Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
Sugawara, Takeyuki Department of Anatomy, Kitasato University School of Medicine
Hatano, Naoya Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
Kanayama, Naoki Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University ORCID Kaken ID publons researchmap
Magari, Masaki Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University
Sakagami, Hiroyuki Department of Anatomy, Kitasato University School of Medicine
Tokumitsu, Hiroshi Applied Cell Biology, Graduate School of Interdisciplinary Science and Engineering in Health Systems, Okayama University ORCID Kaken ID publons researchmap
抄録
BACKGROUND:
Ca2+/calmodulin-dependent protein kinase kinase (CaMKK) is a pivotal activator of CaMKI, CaMKIV and 5'-AMP-activated protein kinase (AMPK), controlling Ca2+-dependent intracellular signaling including various neuronal, metabolic and pathophysiological responses. Recently, we demonstrated that CaMKKβ is feedback phosphorylated at Thr144 by the downstream AMPK, resulting in the conversion of CaMKKβ into Ca2+/CaM-dependent enzyme. However, the regulatory phosphorylation of CaMKKβ at Thr144 in intact cells and in vivo remains unclear.
METHODS:
Anti-phosphoThr144 antibody was used to characterize the site-specific phosphorylation of CaMKKβ in immunoprecipitated samples from mouse cerebellum and in transfected mammalian cells that were treated with various agonists and protein kinase inhibitors. CaMKK activity assay and LC-MS/MS analysis were used for biochemical characterization of phosphorylated CaMKKβ.
RESULTS:
Our data suggest that the phosphorylation of Thr144 in CaMKKβ is rapidly induced by cAMP/cAMP-dependent protein kinase (PKA) signaling in CaMKKβ-transfected HeLa cells, that is physiologically relevant in mouse cerebellum. We confirmed that the catalytic subunit of PKA was capable of directly phosphorylating CaMKKβ at Thr144 in vitro and in transfected cells. In addition, the basal phosphorylation of CaMKKβ at Thr144 in transfected HeLa cells was suppressed by AMPK inhibitor (compound C). PKA-catalyzed phosphorylation reduced the autonomous activity of CaMKKβ in vitro without significant effect on the Ca2+/CaM-dependent activity, resulting in the conversion of CaMKKβ into Ca2+/CaM-dependent enzyme.
CONCLUSION:
cAMP/PKA signaling may confer Ca2+-dependency to the CaMKKβ-mediated signaling pathway through direct phosphorylation of Thr144 in intact cells.
GENERAL SIGNIFICANCE:
Our results suggest a novel cross-talk between cAMP/PKA and Ca2+/CaM/CaMKKβ signaling through regulatory phosphorylation.
キーワード
CaMKK
Calmodulin
Intracellular Ca(2+)
PKA
Phosphorylation
Signal transduction
発行日
2019-01-17
出版物タイトル
Biochimica et Biophysica Acta (BBA) - General Subjects
1863巻
4号
出版者
Elsevier Science
開始ページ
672
終了ページ
680
ISSN
0304-4165
NCID
AA00564679
資料タイプ
学術雑誌論文
言語
English
OAI-PMH Set
岡山大学
論文のバージョン
author
PubMed ID
DOI
Web of Science KeyUT
関連URL
isVersionOf https://doi.org/10.1016/j.bbagen.2018.12.012
Citation
Shota Takabatake, Satomi Ohtsuka, Takeyuki Sugawara, Naoya Hatano, Naoki Kanayama, Masaki Magari, Hiroyuki Sakagami, Hiroshi Tokumitsu, Regulation of Ca2+/calmodulin-dependent protein kinase kinase β by cAMP signaling, Biochimica et Biophysica Acta (BBA) - General Subjects, Volume 1863, Issue 4, 2019, Pages 672-680, ISSN 0304-4165, https://doi.org/10.1016/j.bbagen.2018.12.012.
助成機関名
日本学術振興会
助成番号
18K06113