Hypoxic induction of the regulator of G-protein signalling 4 gene is mediated by the hypoxia-inducible factor pathway

• Published in: PloS one 2012-09, Vol.7 (9), p.e44564-e44564
• Main Authors: Olechnowicz, Sam W Z, Fedele, Anthony O, Peet, Daniel J
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Basic Helix-Loop-Helix Transcription Factors - metabolism; Binding sites; Biology; Cardiomyopathy; Cell lines; Cells (Biology); Dactinomycin - administration & dosage; DNA binding proteins; DNA Primers; Dopamine; Drugs; Endothelial cells; G protein-coupled receptors; Gene expression; Gene Expression Profiling; Genes; Genetic engineering; Genomes; genomics; Guanine nucleotide-binding protein; Hypoxia; Hypoxia - genetics; Hypoxia-inducible factor 1; Hypoxia-inducible factors; Kinases; Mammalian cells; Mental disorders; Narcotics; Neuroblastoma; Neuroblastoma cells; Oligonucleotide Array Sequence Analysis; PC12 Cells; Personal computers; Pheochromocytoma; Polymerase Chain Reaction; Proteins; Rats; Receptors; RGS Proteins - genetics; RNA, Small Interfering; Rodents; Schizophrenia; Signal transduction; Signaling; Transcription; Transcription (Genetics); Transcription factors; Vascular endothelial growth factor
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0044564

The transcriptional response to hypoxia is largely dependent on the Hypoxia Inducible Factors (HIF-1 and HIF-2) in mammalian cells. Many target genes have been characterised for these heterodimeric transcription factors, yet there is evidence that the full range of HIF-regulated genes has not yet been described. We constructed a TetON overexpression system in the rat pheochromocytoma PC-12 cell line to search for novel HIF and hypoxia responsive genes. The Rgs4 gene encodes the Regulator of G-Protein Signalling 4 (RGS4) protein, an inhibitor of signalling from G-protein coupled receptors, and dysregulation of Rgs4 is linked to disease states such as schizophrenia and cardiomyopathy. Rgs4 was found to be responsive to HIF-2α overexpression, hypoxic treatment, and hypoxia mimetic drugs in PC-12 cells. Similar responses were observed in human neuroblastoma cell lines SK-N-SH and SK-N-BE(2)C, but not in endothelial cells, where Rgs4 transcript is readily detected but does not respond to hypoxia. Furthermore, this regulation was found to be dependent on transcription, and occurs in a manner consistent with direct HIF transactivation of Rgs4 transcription. However, no HIF binding site was detectable within 32 kb of the human Rgs4 gene locus, leading to the possibility of regulation by long-distance genomic interactions. Further research into Rgs4 regulation by hypoxia and HIF may result in better understanding of disease states such as schizophrenia, and also shed light on the other roles of HIF yet to be discovered.