Carbon monoxide promotes VEGF expression by increasing HIF-1α protein level via two distinct mechanisms, translational activation and stabilization of HIF-1α protein

Yoon Kyung Choi, Chun Ki Kim, Hansoo Lee, Dooil Jeoung, Kwon Soo Ha, Young-Guen Kwon, Kyu Won Kim, Young Myeong Kim

Research output: Contribution to journalArticle

82 Citations (Scopus)

Abstract

Carbon monoxide (CO) plays a significant role in vascular functions. We here examined the molecular mechanism by which CO regulates HIF-1 (hypoxia-inducible transcription factor-1)-dependent expression of vascular endothelial growth factor (VEGF), which is an important angiogenic factor. We found that astrocytes stimulated with CORM-2 (CO-releasing molecule) promoted angiogenesis by increasing VEGF expression and secretion. CORM-2 also induced HO-1 (hemeoxygenase-1) expression and increased nuclear HIF-1α protein level, without altering its promoter activity and mRNA level. VEGF expression was inhibited by treatment with HIF-1α siRNA and a hemeoxygenase inhibitor, indicating that CO stimulates VEGF expression via up-regulation of HIF-1α protein level, which is partially associated with HO-1 induction. CORM-2 activated the translational regulatory proteins p70S6k and eIF-4E as well as phosphorylating their upstream signal mediators Akt and ERK. These translational signal events and HIF-1α protein level were suppressed by inhibitors of phosphatidylinositol 3-kinase (PI3K), MEK, and mTOR, suggesting that the PI3K/Akt/mTOR and MEK/ERK pathways are involved in a translational increase in HIF-1α. In addition, CORM-2 also increased stability of the HIF-1α protein by suppressing its ubiquitination, without altering the proline hydroxylase-dependent HIF-1α degradation pathway. CORM-2 increased HIF-1α/HSP90α interaction, which is responsible for HIF-1α stabilization, and HSP90-specific inhibitors decreased this interaction, HIF-1α protein level, and VEGF expression. Furthermore, HSP90α knockdown suppressed CORM-2-induced increases in HIF-1α and VEGF protein levels. These results suggest that CO stimulates VEGF production by increasing HIF-1α protein level via two distinct mechanisms, translational stimulation and protein stabilization of HIF-1α.

Original languageEnglish
Pages (from-to)32116-32125
Number of pages10
JournalJournal of Biological Chemistry
Volume285
Issue number42
DOIs
Publication statusPublished - 2010 Oct 15

Fingerprint

Hypoxia-Inducible Factor 1
Carbon Monoxide
Vascular Endothelial Growth Factor A
Transcription Factors
Stabilization
Chemical activation
Proteins
Phosphatidylinositol 3-Kinase
Heme Oxygenase-1
Mitogen-Activated Protein Kinase Kinases
Prolyl Hydroxylases
70-kDa Ribosomal Protein S6 Kinases
MAP Kinase Signaling System
Angiogenesis Inducing Agents
Ubiquitination
Astrocytes
Small Interfering RNA

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Choi, Yoon Kyung ; Kim, Chun Ki ; Lee, Hansoo ; Jeoung, Dooil ; Ha, Kwon Soo ; Kwon, Young-Guen ; Kim, Kyu Won ; Kim, Young Myeong. / Carbon monoxide promotes VEGF expression by increasing HIF-1α protein level via two distinct mechanisms, translational activation and stabilization of HIF-1α protein. In: Journal of Biological Chemistry. 2010 ; Vol. 285, No. 42. pp. 32116-32125.
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Carbon monoxide promotes VEGF expression by increasing HIF-1α protein level via two distinct mechanisms, translational activation and stabilization of HIF-1α protein. / Choi, Yoon Kyung; Kim, Chun Ki; Lee, Hansoo; Jeoung, Dooil; Ha, Kwon Soo; Kwon, Young-Guen; Kim, Kyu Won; Kim, Young Myeong.

In: Journal of Biological Chemistry, Vol. 285, No. 42, 15.10.2010, p. 32116-32125.

Research output: Contribution to journalArticle

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T1 - Carbon monoxide promotes VEGF expression by increasing HIF-1α protein level via two distinct mechanisms, translational activation and stabilization of HIF-1α protein

AU - Choi, Yoon Kyung

AU - Kim, Chun Ki

AU - Lee, Hansoo

AU - Jeoung, Dooil

AU - Ha, Kwon Soo

AU - Kwon, Young-Guen

AU - Kim, Kyu Won

AU - Kim, Young Myeong

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N2 - Carbon monoxide (CO) plays a significant role in vascular functions. We here examined the molecular mechanism by which CO regulates HIF-1 (hypoxia-inducible transcription factor-1)-dependent expression of vascular endothelial growth factor (VEGF), which is an important angiogenic factor. We found that astrocytes stimulated with CORM-2 (CO-releasing molecule) promoted angiogenesis by increasing VEGF expression and secretion. CORM-2 also induced HO-1 (hemeoxygenase-1) expression and increased nuclear HIF-1α protein level, without altering its promoter activity and mRNA level. VEGF expression was inhibited by treatment with HIF-1α siRNA and a hemeoxygenase inhibitor, indicating that CO stimulates VEGF expression via up-regulation of HIF-1α protein level, which is partially associated with HO-1 induction. CORM-2 activated the translational regulatory proteins p70S6k and eIF-4E as well as phosphorylating their upstream signal mediators Akt and ERK. These translational signal events and HIF-1α protein level were suppressed by inhibitors of phosphatidylinositol 3-kinase (PI3K), MEK, and mTOR, suggesting that the PI3K/Akt/mTOR and MEK/ERK pathways are involved in a translational increase in HIF-1α. In addition, CORM-2 also increased stability of the HIF-1α protein by suppressing its ubiquitination, without altering the proline hydroxylase-dependent HIF-1α degradation pathway. CORM-2 increased HIF-1α/HSP90α interaction, which is responsible for HIF-1α stabilization, and HSP90-specific inhibitors decreased this interaction, HIF-1α protein level, and VEGF expression. Furthermore, HSP90α knockdown suppressed CORM-2-induced increases in HIF-1α and VEGF protein levels. These results suggest that CO stimulates VEGF production by increasing HIF-1α protein level via two distinct mechanisms, translational stimulation and protein stabilization of HIF-1α.

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