title: AT1 receptor induced alterations in histone H2A reveal novel insights into GPCR control of chromatin remodeling.
abstract: Chronic activation of angiotensin II (AngII) type 1 receptor (AT(1)R), a prototypical G protein-coupled receptor (GPCR) induces gene regulatory stress which is responsible for phenotypic modulation of target cells. The AT(1)R-selective drugs reverse the gene regulatory stress in various cardiovascular diseases. However, the molecular mechanisms are not clear. We speculate that activation states of AT(1)R modify the composition of histone isoforms and post-translational modifications (PTM), thereby alter the structure-function dynamics of chromatin. We combined total histone isolation, FPLC separation, and mass spectrometry techniques to analyze histone H2A in HEK293 cells with and without AT(1)R activation. We have identified eight isoforms: H2AA, H2AG, H2AM, H2AO, H2AQ, Q96QV6, H2AC and H2AL. The isoforms, H2AA, H2AC and H2AQ were methylated and H2AC was phosphorylated. The relative abundance of specific H2A isoforms and PTMs were further analyzed in relationship to the activation states of AT(1)R by immunochemical studies. Within 2 hr, the isoforms, H2AA/O exchanged with H2AM. The monomethylated H2AC increased rapidly and the phosphorylated H2AC decreased, thus suggesting that enhanced H2AC methylation is coupled to Ser1p dephosphorylation. We show that H2A125Kme1 promotes interaction with the heterochromatin associated protein, HP1α. These specific changes in H2A are reversed by treatment with the AT(1)R specific inhibitor losartan. Our analysis provides a first step towards an awareness of histone code regulation by GPCRs.
authors.affiliation: Department of Molecular Cardiology, Cleveland Clinic Foundation, Cleveland, Ohio, United States of America.
authors.forename: Rajaganapathi
authors.initials: R
authors.lastname: Jagannathan
authors.forename: Suma
authors.initials: S
authors.lastname: Kaveti
authors.forename: Russell W
authors.initials: RW
authors.lastname: Desnoyer
authors.forename: Belinda
authors.initials: B
authors.lastname: Willard
authors.forename: Michael
authors.initials: M
authors.lastname: Kinter
authors.forename: Sadashiva S
authors.initials: SS
authors.lastname: Karnik
mesh_terms: D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly; D000090266:Chromobox Protein Homolog 5; D057809:HEK293 Cells; D006570:Heterochromatin / Q000235:genetics / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008745:Methylation; D010766:Phosphorylation; D044140:Receptor, Angiotensin, Type 1 / Q000235:genetics / Q000378:metabolism*
chemical_list: C000715594:CBX5 protein, human; D002843:Chromatin; D006570:Heterochromatin; D006657:Histones; D044140:Receptor, Angiotensin, Type 1; D000090266:Chromobox Protein Homolog 5