title: Transcriptional regulation of the novel monoamine oxidase renalase: Crucial roles of transcription factors Sp1, STAT3, and ZBP89.
abstract: Renalase, a novel monoamine oxidase, is emerging as an important regulator of cardiovascular, metabolic, and renal diseases. However, the mechanism of transcriptional regulation of this enzyme remains largely unknown. We undertook a systematic analysis of the renalase gene to identify regulatory promoter elements and transcription factors. Computational analysis coupled with transfection of human renalase promoter/luciferase reporter plasmids (5'-promoter-deletion constructs) into various cell types (HEK-293, IMR32, and HepG2) identified two crucial promoter domains at base pairs -485 to -399 and -252 to -150. Electrophoretic mobility shift assays using renalase promoter oligonucleotides with and without potential binding sites for transcription factors Sp1, STAT3, and ZBP89 displayed formation of specific complexes with HEK-293 nuclear proteins. Consistently, overexpression of Sp1, STAT3, and ZBP89 augmented renalase promoter activity; additionally, siRNA-mediated downregulation of Sp1, STAT3, and ZBP89 reduced the level of endogenous renalase transcription as well as the transfected renalase promoter activity. In addition, chromatin immunoprecipitation assays showed in vivo interactions of these transcription factors with renalase promoter. Interestingly, renalase promoter activity was augmented by nicotine and catecholamines; while Sp1 and STAT3 synergistically activated the nicotine-induced effect, Sp1 appeared to enhance epinephrine-evoked renalase transcription. Moreover, renalase transcript levels in mouse models of human essential hypertension were concomitantly associated with endogenous STAT3 and ZBP89 levels, suggesting crucial roles for these transcription factors in regulating renalase gene expression in cardiovascular pathological conditions.
authors.affiliation: Cardiovascular Genetics Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras , Chennai 600036, India.
authors.forename: Parshuram J
authors.initials: PJ
authors.lastname: Sonawane
authors.forename: Vinayak
authors.initials: V
authors.lastname: Gupta
authors.forename: Binu K
authors.initials: BK
authors.lastname: Sasi
authors.forename: Ananthamohan
authors.initials: A
authors.lastname: Kalyani
authors.forename: Bhargavi
authors.initials: B
authors.lastname: Natarajan
authors.forename: Abrar A
authors.initials: AA
authors.lastname: Khan
authors.forename: Bhavani S
authors.initials: BS
authors.lastname: Sahu
authors.forename: Nitish R
authors.initials: NR
authors.lastname: Mahapatra
mesh_terms: D000818:Animals; D001483:Base Sequence; D016384:Consensus Sequence; D004268:DNA-Binding Proteins / Q000502:physiology*; D000075222:Essential Hypertension; D015971:Gene Expression Regulation, Enzymologic; D057809:HEK293 Cells; D056945:Hep G2 Cells; D006801:Humans; D006973:Hypertension / Q000201:enzymology / Q000235:genetics; D008297:Male; D008815:Mice, Inbred Strains; D008995:Monoamine Oxidase / Q000235:genetics* / Q000378:metabolism; D009538:Nicotine / Q000494:pharmacology; D018722:Nicotinic Agonists / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D050796:STAT3 Transcription Factor / Q000502:physiology*; D016329:Sp1 Transcription Factor / Q000502:physiology*; D014157:Transcription Factors / Q000502:physiology*; D014158:Transcription, Genetic; D015533:Transcriptional Activation
chemical_list: D004268:DNA-Binding Proteins; D018722:Nicotinic Agonists; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D016329:Sp1 Transcription Factor; C108209:SP1 protein, human; D014157:Transcription Factors; C110951:ZNF148 protein, human; D009538:Nicotine; D008995:Monoamine Oxidase; C521557:renalase