Hypoxia-inducible factors regulate human and rat cystathionine {Sharp S}-synthase gene expression

Increased catalytic activity of cystathionine ß-synthase (CBS) was recently shown to mediate vasodilation of the cerebral microcirculation, which is initiated within minutes after the onset of acute hypoxia. To test whether chronic hypoxia was a stimulus for increased CBS expression, U87-MG human glioblastoma and PC12 rat pheochromocytoma cells were exposed to 1% or 20% O2 for 24 to 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 short hairpin RNA targeting hypoxia-inducible factor (HIF) 1alpha or 2alpha. Exposure of rats to hypobaric hypoxia (0.35 atm) for 3 d 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 chromatin immunoprecipitation assays. A 49-bp human sequence, which encompassed an inverted repeat of the core HIF binding site, functioned as a hypoxia response element in luciferase reporter transcription assays. Thus, HIFs mediate tissue-specific CBS expression, which may augment cerebral vasodilation as an adaptive response to chronic hypoxia.
Source: BJ Gene - Category: Biochemistry Authors: Tags: BJ Gene Source Type: research