PMID- 27865915 OWN - NLM STAT- MEDLINE DCOM- 20170531 LR - 20190221 IS - 1095-8584 (Electronic) IS - 0022-2828 (Linking) VI - 102 DP - 2017 Jan TI - Functional promoter polymorphisms direct the expression of cystathionine gamma-lyase gene in mouse models of essential hypertension. PG - 61-73 LID - S0022-2828(16)30409-6 [pii] LID - 10.1016/j.yjmcc.2016.11.005 [doi] AB - Despite the well-known role of cystathionine gamma-lyase (Cth) in cardiovascular pathophysiology, transcriptional regulation of Cth remains incompletely understood. Sequencing of the Cth promoter region in mouse models of genetic/essential hypertension (viz. Blood Pressure High [BPH], Blood Pressure Low [BPL] and Blood Pressure Normal [BPN] mice) identified several genetic variations. Transient transfections of BPH/BPL-Cth promoter-reporter plasmids into various cell types revealed higher promoter activity of BPL-Cth than that of BPH-Cth. Corroboratively, endogenous Cth mRNA levels in kidney and liver tissues were also elevated in BPL mice. Computational analysis of the polymorphic Cth promoter region predicted differential binding affinity of c-Rel, HOXA3 and IRF1 with BPL/BPH-Cth promoter domains. Over-expression of c-Rel/HOXA3/IRF1 modulated BPL/BPH-Cth promoter activities in a consistent manner. Gel shift assays using BPH/BPL-Cth-promoter oligonucleotides with/without binding sites for c-Rel/HOXA3/IRF1 displayed formation of specific complexes with c-Rel/HOXA3/IRF1; addition of antibodies to reaction mixtures resulted in supershifts/inhibition of Cth promoter-transcription factor complexes. Furthermore, chromatin immunoprecipitation (ChIP) assays proved differential binding of c-Rel, HOXA3 and IRF1 with the polymorphic promoter region of BPL/BPH-Cth. Tumor necrosis factor-alpha (TNF-alpha) reduced the activities of BPL/BPH-Cth promoters to different extents that were further declined by ectopic expression of IRF1; on the other hand, siRNA-mediated down-regulation of IRF1 rescued the TNF-alpha-mediated suppression of the BPL/BPH-Cth promoter activities. In corroboration, ChIP analysis revealed enhanced binding of IRF1 with BPH/BPL-Cth promoter following TNF-alpha treatment. BPL/BPH-Cth promoter activity was diminished upon exposure of hepatocytes and cardiomyoblasts to ischemia-like pathological condition due to reduced binding of c-Rel with BPL/BPH-Cth-promoter. Taken together, this study reveals the molecular basis for the differential expression of Cth in mouse models of essential hypertension under basal and pathophysiological conditions. CI - Copyright A(c) 2016 Elsevier Ltd. All rights reserved. FAU - Gupta, Vinayak AU - Gupta V AD - Cardiovascular Genetics Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, India. FAU - Kapopara, Piyushkumar R AU - Kapopara PR AD - Cardiovascular Genetics Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, India. FAU - Khan, Abrar A AU - Khan AA AD - Cardiovascular Genetics Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, India. FAU - Arige, Vikas AU - Arige V AD - Cardiovascular Genetics Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, India. FAU - Subramanian, Lakshmi AU - Subramanian L AD - Cardiovascular Genetics Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, India. FAU - Sonawane, Parshuram J AU - Sonawane PJ AD - Cardiovascular Genetics Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, India. FAU - Sasi, Binu K AU - Sasi BK AD - Cardiovascular Genetics Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, India. FAU - Mahapatra, Nitish R AU - Mahapatra NR AD - Cardiovascular Genetics Laboratory, Department of Biotechnology, Bhupat and Jyoti Mehta School of Biosciences, Indian Institute of Technology Madras, Chennai 600036, India. Electronic address: nmahapatra@iitm.ac.in. LA - eng PT - Journal Article DEP - 20161116 PL - England TA - J Mol Cell Cardiol JT - Journal of molecular and cellular cardiology JID - 0262322 RN - 0 (Transcription Factors) RN - 0 (Tumor Necrosis Factor-alpha) RN - EC 4.4.1.1 (Cystathionine gamma-Lyase) SB - IM MH - Animals MH - Base Sequence MH - Binding Sites MH - Blood Pressure MH - Chromosome Mapping MH - Computational Biology/methods MH - Cystathionine gamma-Lyase/*genetics MH - Disease Models, Animal MH - Essential Hypertension MH - *Gene Expression Regulation MH - Genomics/methods MH - Hypertension/*genetics/*physiopathology MH - Mice MH - Nucleotide Motifs MH - Organ Specificity/genetics MH - *Polymorphism, Genetic MH - *Promoter Regions, Genetic MH - Protein Binding MH - Quantitative Trait Loci MH - Rats MH - Transcription Factors/metabolism MH - Transcription, Genetic MH - Tumor Necrosis Factor-alpha/metabolism OTO - NOTNLM OT - Cystathionine gamma-lyase promoter OT - Gene regulation OT - Ischemia OT - Transcription factor OT - Tumor necrosis factor-alpha EDAT- 2016/11/21 06:00 MHDA- 2017/06/01 06:00 CRDT- 2016/11/21 06:00 PHST- 2016/02/03 00:00 [received] PHST- 2016/10/21 00:00 [revised] PHST- 2016/11/11 00:00 [accepted] PHST- 2016/11/21 06:00 [pubmed] PHST- 2017/06/01 06:00 [medline] PHST- 2016/11/21 06:00 [entrez] AID - S0022-2828(16)30409-6 [pii] AID - 10.1016/j.yjmcc.2016.11.005 [doi] PST - ppublish SO - J Mol Cell Cardiol. 2017 Jan;102:61-73. doi: 10.1016/j.yjmcc.2016.11.005. Epub 2016 Nov 16.