Tensile strain-induced Ets-2 phosphorylation by CaMKII and the homeostasis of cranial sutures

Jack C Yu, Jung Ren Chen, Chao Hsiung Lin, Guigen Zhang, Poh Sang Lam, Karl H. Wenger, Farid B. Mozaffari, Shun Te Huang, James L. Borke

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background: Mechanotransduction underpins the homeostasis of musculoskeletal tissues, including cranial sutures. Intracellular calcium, [Ca 2+]ic, and protein phosphorylation are two intermediate variables in signal relay during mechanotransduction. This project establishes a chain of cause and effect, linking cellular strain to substrate phosphorylation, and identifies the agent and target sites of phosphorylation. Methods: Cyclic tensile force (0.5 N at 1 Hz) was applied to 1-day-old rat sagittal sutures. [Ca]ic was measured by FURA-2. Ets-2 phosphorylation by CaMKII was tested using Western blot autoradiography. Peptide array was constructed to determine the precise sites of phosphorylation. The results were confirmed with mass spectroscopy and Western blots using phospho-specific antibodies. Results: [Ca 2+]ic increased rapidly in response to tensile stress. In the presence of Ca, CaMKII caused Ets-2 phosphorylation. Of the three possible sites for phosphorylation of Ets-2 by CaMKII, RVPS, FESF, RLSS, Serine 246, 310, and 313 were the targets. Furthermore, the contiguous sequence modified this effect. Mass spectroscopy showed 80 Da (molecular weight of phosphate group, -PO3) right shifts consistent with phosphorylation. There was cytosolic translocation of Ets-2 on tensile deformation of suture cells. CaMKII binding of Ets-2 occurred within 30 minutes after the onset of tensile strain. Conclusions: Cranial suture cells can respond to tensile forces by increasing [Ca 2+]ic, which causes CaMKII to phosphorylate Ets-2, thus altering Ets-2 binding to its downstream promoters. Of note, Ets-2 is at the intersection of three key pathways important in craniosynostosis: fibroblast growth factor-2, transforming growth factor-β, and mechanotransduction.

Original languageEnglish (US)
JournalPlastic and reconstructive surgery
Volume123
Issue numberSUPPL.2S
DOIs
StatePublished - Sep 18 2009

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Cranial Sutures
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Homeostasis
Phosphorylation
Sutures
Mass Spectrometry
Western Blotting
Phospho-Specific Antibodies
Craniosynostoses
Transforming Growth Factors
Fibroblast Growth Factor 2
Autoradiography
Serine
Molecular Weight
Phosphates
Calcium
Peptides

ASJC Scopus subject areas

  • Surgery

Cite this

Tensile strain-induced Ets-2 phosphorylation by CaMKII and the homeostasis of cranial sutures. / Yu, Jack C; Chen, Jung Ren; Lin, Chao Hsiung; Zhang, Guigen; Lam, Poh Sang; Wenger, Karl H.; Mozaffari, Farid B.; Huang, Shun Te; Borke, James L.

In: Plastic and reconstructive surgery, Vol. 123, No. SUPPL.2S, 18.09.2009.

Research output: Contribution to journalArticle

Yu, JC, Chen, JR, Lin, CH, Zhang, G, Lam, PS, Wenger, KH, Mozaffari, FB, Huang, ST & Borke, JL 2009, 'Tensile strain-induced Ets-2 phosphorylation by CaMKII and the homeostasis of cranial sutures', Plastic and reconstructive surgery, vol. 123, no. SUPPL.2S. https://doi.org/10.1097/PRS.0b013e318191c029
Yu, Jack C ; Chen, Jung Ren ; Lin, Chao Hsiung ; Zhang, Guigen ; Lam, Poh Sang ; Wenger, Karl H. ; Mozaffari, Farid B. ; Huang, Shun Te ; Borke, James L. / Tensile strain-induced Ets-2 phosphorylation by CaMKII and the homeostasis of cranial sutures. In: Plastic and reconstructive surgery. 2009 ; Vol. 123, No. SUPPL.2S.
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T1 - Tensile strain-induced Ets-2 phosphorylation by CaMKII and the homeostasis of cranial sutures

AU - Yu, Jack C

AU - Chen, Jung Ren

AU - Lin, Chao Hsiung

AU - Zhang, Guigen

AU - Lam, Poh Sang

AU - Wenger, Karl H.

AU - Mozaffari, Farid B.

AU - Huang, Shun Te

AU - Borke, James L.

PY - 2009/9/18

Y1 - 2009/9/18

N2 - Background: Mechanotransduction underpins the homeostasis of musculoskeletal tissues, including cranial sutures. Intracellular calcium, [Ca 2+]ic, and protein phosphorylation are two intermediate variables in signal relay during mechanotransduction. This project establishes a chain of cause and effect, linking cellular strain to substrate phosphorylation, and identifies the agent and target sites of phosphorylation. Methods: Cyclic tensile force (0.5 N at 1 Hz) was applied to 1-day-old rat sagittal sutures. [Ca]ic was measured by FURA-2. Ets-2 phosphorylation by CaMKII was tested using Western blot autoradiography. Peptide array was constructed to determine the precise sites of phosphorylation. The results were confirmed with mass spectroscopy and Western blots using phospho-specific antibodies. Results: [Ca 2+]ic increased rapidly in response to tensile stress. In the presence of Ca, CaMKII caused Ets-2 phosphorylation. Of the three possible sites for phosphorylation of Ets-2 by CaMKII, RVPS, FESF, RLSS, Serine 246, 310, and 313 were the targets. Furthermore, the contiguous sequence modified this effect. Mass spectroscopy showed 80 Da (molecular weight of phosphate group, -PO3) right shifts consistent with phosphorylation. There was cytosolic translocation of Ets-2 on tensile deformation of suture cells. CaMKII binding of Ets-2 occurred within 30 minutes after the onset of tensile strain. Conclusions: Cranial suture cells can respond to tensile forces by increasing [Ca 2+]ic, which causes CaMKII to phosphorylate Ets-2, thus altering Ets-2 binding to its downstream promoters. Of note, Ets-2 is at the intersection of three key pathways important in craniosynostosis: fibroblast growth factor-2, transforming growth factor-β, and mechanotransduction.

AB - Background: Mechanotransduction underpins the homeostasis of musculoskeletal tissues, including cranial sutures. Intracellular calcium, [Ca 2+]ic, and protein phosphorylation are two intermediate variables in signal relay during mechanotransduction. This project establishes a chain of cause and effect, linking cellular strain to substrate phosphorylation, and identifies the agent and target sites of phosphorylation. Methods: Cyclic tensile force (0.5 N at 1 Hz) was applied to 1-day-old rat sagittal sutures. [Ca]ic was measured by FURA-2. Ets-2 phosphorylation by CaMKII was tested using Western blot autoradiography. Peptide array was constructed to determine the precise sites of phosphorylation. The results were confirmed with mass spectroscopy and Western blots using phospho-specific antibodies. Results: [Ca 2+]ic increased rapidly in response to tensile stress. In the presence of Ca, CaMKII caused Ets-2 phosphorylation. Of the three possible sites for phosphorylation of Ets-2 by CaMKII, RVPS, FESF, RLSS, Serine 246, 310, and 313 were the targets. Furthermore, the contiguous sequence modified this effect. Mass spectroscopy showed 80 Da (molecular weight of phosphate group, -PO3) right shifts consistent with phosphorylation. There was cytosolic translocation of Ets-2 on tensile deformation of suture cells. CaMKII binding of Ets-2 occurred within 30 minutes after the onset of tensile strain. Conclusions: Cranial suture cells can respond to tensile forces by increasing [Ca 2+]ic, which causes CaMKII to phosphorylate Ets-2, thus altering Ets-2 binding to its downstream promoters. Of note, Ets-2 is at the intersection of three key pathways important in craniosynostosis: fibroblast growth factor-2, transforming growth factor-β, and mechanotransduction.

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