Hypoxia-inducible factors regulate human and rat cystathionine â-synthase gene expression

Naoharu Takano, Ying Jie Peng, Ganesh K. Kumar, Weibo Luo, Hongxia Hu, Larissa A. Shimoda, Makoto Suematsu, Nanduri R. Prabhakar, Gregg L. Semenza

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Increased catalytic activity of CBS (cystathionine β-synthase) was recently shown to mediate vasodilation of the cerebral microcirculation, which is initiated within minutes of the onset of acute hypoxia. To test whether chronic hypoxia was a stimulus for increased CBS expression, U87-MG human glioblastoma and PC12 rat phaeochromocytoma cells were exposed to 1% or 20%O2 for 24-72 h. CBS mRNA and protein expression were increased in hypoxic cells. Hypoxic induction of CBS expression was abrogated in cells transfected with vector encoding shRNA targeting HIF (hypoxia-inducible factor) 1α or 2α. Exposure of rats to hypobaric hypoxia (0.35 atm; 1 atm = 101.325 kPa) for 3 days induced increased CBS mRNA, protein and catalytic activity in the cerebral cortex and cerebellum, which was blocked by administration of the HIF inhibitor digoxin. HIF-binding sites, located 0.8 and 1.2 kb 5 to the transcription start site of the human CBS and rat Cbs genes respectively, were identified by ChIP assays. A 49-bp human sequence, which encompassed an inverted repeat of the core HIF-binding site, functioned as a hypoxiaresponse element in luciferase reporter transcription assays. Thus HIFsmediate tissue-specificCBSexpression, whichmay augment cerebral vasodilation as an adaptive response to chronic hypoxia.

Original languageEnglish
Pages (from-to)203-211
Number of pages9
JournalBiochemical Journal
Volume458
Issue number2
DOIs
Publication statusPublished - 2014 Mar 1

Fingerprint

Cystathionine
Human engineering
Gene expression
Rats
Gene Expression
Assays
Catalyst activity
Binding Sites
Microcirculation
Hypoxia-Inducible Factor 1
Messenger RNA
Transcription Initiation Site
Digoxin
Transcription
Luciferases
Small Interfering RNA
Vasodilation
Proteins
Genes
Cells

Keywords

  • Cerebral circulation
  • Cystathionine β-synthase (CBS)
  • Hydrogen sulfide
  • Hypoxia-inducible factor (HIF)
  • Oxygen

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Takano, N., Peng, Y. J., Kumar, G. K., Luo, W., Hu, H., Shimoda, L. A., ... Semenza, G. L. (2014). Hypoxia-inducible factors regulate human and rat cystathionine â-synthase gene expression. Biochemical Journal, 458(2), 203-211. https://doi.org/10.1042/BJ20131350

Hypoxia-inducible factors regulate human and rat cystathionine â-synthase gene expression. / Takano, Naoharu; Peng, Ying Jie; Kumar, Ganesh K.; Luo, Weibo; Hu, Hongxia; Shimoda, Larissa A.; Suematsu, Makoto; Prabhakar, Nanduri R.; Semenza, Gregg L.

In: Biochemical Journal, Vol. 458, No. 2, 01.03.2014, p. 203-211.

Research output: Contribution to journalArticle

Takano, N, Peng, YJ, Kumar, GK, Luo, W, Hu, H, Shimoda, LA, Suematsu, M, Prabhakar, NR & Semenza, GL 2014, 'Hypoxia-inducible factors regulate human and rat cystathionine â-synthase gene expression', Biochemical Journal, vol. 458, no. 2, pp. 203-211. https://doi.org/10.1042/BJ20131350
Takano, Naoharu ; Peng, Ying Jie ; Kumar, Ganesh K. ; Luo, Weibo ; Hu, Hongxia ; Shimoda, Larissa A. ; Suematsu, Makoto ; Prabhakar, Nanduri R. ; Semenza, Gregg L. / Hypoxia-inducible factors regulate human and rat cystathionine â-synthase gene expression. In: Biochemical Journal. 2014 ; Vol. 458, No. 2. pp. 203-211.
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