Induction of Hypoxia-inducible Factor 1 Activity by Muscarinic Acetylcholine Receptor Signaling

• Published in: The Journal of biological chemistry 2004-10, Vol.279 (40), p.41521-41528
• Main Authors: Hirota, Kiichi, Fukuda, Ryo, Takabuchi, Satoshi, Kizaka-Kondoh, Shinae, Adachi, Takehiko, Fukuda, Kazuhiko, Semenza, Gregg L
• Format: Article
• Language: English
• Subjects: Cell Line; Gene Expression Regulation; Humans; Hydroxylation; Hypoxia-Inducible Factor 1, alpha Subunit; Mitogen-Activated Protein Kinases - metabolism; Phosphatidylinositol 3-Kinases - metabolism; Receptor Protein-Tyrosine Kinases - metabolism; Receptor, Muscarinic M1 - genetics; Receptor, Muscarinic M1 - physiology; Receptor, Muscarinic M3 - genetics; Receptor, Muscarinic M3 - physiology; Receptors, Muscarinic - genetics; Receptors, Muscarinic - physiology; Signal Transduction; Transcription Factors - biosynthesis; Transcription Factors - metabolism; Transcription, Genetic; Transfection
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M405164200

Hypoxia-inducible factor-1 (HIF-1) is a master regulator of cellular adaptive responses to hypoxia. Levels of the HIF-1α subunit increase under hypoxic conditions. Exposure of cells to growth factors, prostaglandin, and certain nitric oxide donors also induces HIF-1α expression under non-hypoxic conditions. We demonstrate that muscarinic acetylcholine signals induce HIF-1α expression and transcriptional activity in a receptor subtype-specific manner using HEK293 cells transiently overexpressing each of M1-M4 muscarinic acetylcholine receptors. The muscarinic signaling pathways inhibited HIF-1α hydroxylation and degradation and induced HIF-1α protein synthesis that was confirmed by pulse labeling studies. Muscarinic signal-induced HIF-1α protein and HIF-1-dependent gene expression were blocked by treating cells with inhibitors of phosphatidylinositol 3-kinase, MAP kinase kinase, or tyrosine kinase signaling pathways. Dominant-negative forms of Ras and/or Rac-1 significantly suppressed HIF-1 activation by muscarinic signaling. Signaling via M1- and M3- but not M2- or M4-AchRs promote accumulation and transcriptional activation of HIF-1α. We conclude that muscarinic acetylcholine signals activate HIF-1 by both stabilization and synthesis of HIF-1α and by inducing the transcriptional activity of HIF-1α.