Nuclear cytoglobin associates with HMGB2 and regulates DNA damage and genome-wide transcriptional output in the vasculature

Identifying novel regulators of vascular smooth muscle cell function is necessary to further understand cardiovascular diseases. We previously identified cytoglobin, a hemoglobin homolog, with myogenic and cytoprotective roles in the vasculature. The specific mechanism of action of cytoglobin is unclear but does not seem to be related to oxygen transport or storage like hemoglobin. Herein, transcriptomic profiling of injured carotid arteries in cytoglobin global knockout mice revealed that cytoglobin deletion accelerated the loss of contractile genes and increased DNA damage. Overall, we show that cytoglobin is actively translocated into the nucleus of vascular smooth muscle cells through a redox signal driven by NOX4. We demonstrate that nuclear cytoglobin heterodimerizes with the non-histone chromatin structural protein HMGB2. Our results are consistent with a previously unknown function by which a non-erythrocytic hemoglobin inhibits DNA damage and regulates gene programs in the vasculature by modulating the genome-wide binding of HMGB2.

[1]  J. Coon,et al.  A role for cytoglobin in regulating intracellular hydrogen peroxide and redox signals in the vasculature , 2023, bioRxiv.

[2]  M. Bustin,et al.  Epigenetic regulation of white adipose tissue plasticity and energy metabolism by nucleosome binding HMGN proteins , 2022, Nature Communications.

[3]  W. Abplanalp,et al.  Nox4 promotes endothelial differentiation through chromatin remodeling , 2022, Redox biology.

[4]  J. Ayté,et al.  The Mitochondria-to-Cytosol H2O2 Gradient Is Caused by Peroxiredoxin-Dependent Cytosolic Scavenging , 2021, Antioxidants.

[5]  Evgenia Shishkova,et al.  Argonaut: A Web Platform for Collaborative Multi-omic Data Visualization and Exploration , 2020, Patterns.

[6]  M. Bianchi,et al.  Nucleosomes effectively shield DNA from radiation damage in living cells , 2020, Nucleic acids research.

[7]  Natalie M. Mishina,et al.  Ultrasensitive Genetically Encoded Indicator for Hydrogen Peroxide Identifies Roles for the Oxidant in Cell Migration and Mitochondrial Function. , 2020, Cell metabolism.

[8]  D. Jourd’heuil,et al.  Emerging perspectives on cytoglobin, beyond NO dioxygenase and peroxidase , 2020, Redox biology.

[9]  J. Zweier,et al.  Regulation of Nitric Oxide Metabolism and Vascular Tone by Cytoglobin. , 2019, Antioxidants & redox signaling.

[10]  Kejiao Li,et al.  A Genetic Map of the Response to DNA Damage in Human Cells , 2019, Cell.

[11]  Young Hun Kim,et al.  Peroxiredoxin-mediated disulfide bond formation is required for nucleocytoplasmic translocation and secretion of HMGB1 in response to inflammatory stimuli , 2019, Redox biology.

[12]  M. Westphall,et al.  Ultra-High Pressure (>30,000 psi) Packing of Capillary Columns Enhancing Depth of Shotgun Proteomic Analyses. , 2018, Analytical chemistry.

[13]  K. Rippe,et al.  HMGB2 Loss upon Senescence Entry Disrupts Genomic Organization and Induces CTCF Clustering across Cell Types. , 2018, Molecular cell.

[14]  Matteo Pellegrini,et al.  High-Resolution Mapping of Chromatin Conformation in Cardiac Myocytes Reveals Structural Remodeling of the Epigenome in Heart Failure , 2017, Circulation.

[15]  Shunlin Qu,et al.  Cytoglobin Promotes Cardiac Progenitor Cell Survival against Oxidative Stress via the Upregulation of the NFκB/iNOS Signal Pathway and Nitric Oxide Production , 2017, Scientific Reports.

[16]  M. Gladwin,et al.  Efficient Reduction of Vertebrate Cytoglobins by the Cytochrome b5/Cytochrome b5 Reductase/NADH System. , 2017, Biochemistry.

[17]  Xiaoping Liu,et al.  Cytoglobin regulates blood pressure and vascular tone through nitric oxide metabolism in the vascular wall , 2017, Nature Communications.

[18]  P. Gallagher,et al.  Inflammation and vascular smooth muscle cell dedifferentiation following carotid artery ligation. , 2017, Physiological genomics.

[19]  Benjamin G. Bitler,et al.  HMGB2 orchestrates the chromatin landscape of senescence-associated secretory phenotype gene loci , 2016, The Journal of cell biology.

[20]  Christoph D. Rau,et al.  Reciprocal Regulation of the Cardiac Epigenome by Chromatin Structural Proteins Hmgb and Ctcf , 2016, The Journal of Biological Chemistry.

[21]  K. Yoshizato,et al.  Absence of cytoglobin promotes multiple organ abnormalities in aged mice , 2016, Scientific Reports.

[22]  Minyoung Lee,et al.  Redox-Sensitive Regulation of Myocardin-Related Transcription Factor (MRTF-A) Phosphorylation via Palladin in Vascular Smooth Muscle Cell Differentiation Marker Gene Expression , 2016, PloS one.

[23]  M. Bennett,et al.  Vascular Smooth Muscle Cells in Atherosclerosis. , 2016, Circulation research.

[24]  G. Owens,et al.  Epigenetic Control of Smooth Muscle Cell Identity and Lineage Memory. , 2015, Arteriosclerosis, thrombosis, and vascular biology.

[25]  Diana C. Canseco,et al.  Cytoglobin modulates myogenic progenitor cell viability and muscle regeneration , 2013, Proceedings of the National Academy of Sciences.

[26]  Andreas Krämer,et al.  Causal analysis approaches in Ingenuity Pathway Analysis , 2013, Bioinform..

[27]  M. Rudnicki,et al.  Oct4 Interaction with Hmgb2 Regulates Akt Signaling and Pluripotency , 2013, Stem cells.

[28]  Xue-chuan Shi,et al.  Mechanisms of Neuroprotection from Hypoxia-Ischemia (HI) Brain Injury by Up-regulation of Cytoglobin (CYGB) in a Neonatal Rat Model* , 2013, The Journal of Biological Chemistry.

[29]  A. Nakamura,et al.  Use of the γ-H2AX assay to monitor DNA damage and repair in translational cancer research. , 2012, Cancer letters.

[30]  N. Friedman,et al.  Substantial Histone Reduction Modulates Genomewide Nucleosomal Occupancy and Global Transcriptional Output , 2011, PLoS biology.

[31]  Guoqing Hu,et al.  Repression of Smooth Muscle Differentiation by a Novel High Mobility Group Box-containing Protein, HMG2L1* , 2010, The Journal of Biological Chemistry.

[32]  Xingzhi Xu,et al.  Protein Phosphatase 6 Interacts with the DNA-Dependent Protein Kinase Catalytic Subunit and Dephosphorylates γ-H2AX , 2010, Molecular and Cellular Biology.

[33]  N. Krynetskaia,et al.  Chromatin-associated proteins HMGB1/2 and PDIA3 trigger cellular response to chemotherapy-induced DNA damage , 2009, Molecular Cancer Therapeutics.

[34]  D. Jourd’heuil,et al.  Cytoglobin Is Expressed in the Vasculature and Regulates Cell Respiration and Proliferation via Nitric Oxide Dioxygenation* , 2009, Journal of Biological Chemistry.

[35]  M. Mann,et al.  MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification , 2008, Nature Biotechnology.

[36]  A. Schechter.  Hemoglobin research and the origins of molecular medicine. , 2008, Blood.

[37]  M. Graham,et al.  Cellular protection from oxidative DNA damage by over-expression of the novel globin cytoglobin in vitro. , 2008, Mutagenesis.

[38]  M. Hargrove,et al.  NO Dioxygenase Activity in Hemoglobins Is Ubiquitous In Vitro, but Limited by Reduction In Vivo , 2008, PloS one.

[39]  D. Sorescu,et al.  Nox4 Is Required for Maintenance of the Differentiated Vascular Smooth Muscle Cell Phenotype , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[40]  G. Owens,et al.  Molecular regulation of vascular smooth muscle cell differentiation in development and disease. , 2004, Physiological reviews.

[41]  E. Nevo,et al.  Cytoglobin Is a Respiratory Protein in Connective Tissue and Neurons, Which Is Up-regulated by Hypoxia* , 2004, Journal of Biological Chemistry.

[42]  Alessandra Pesce,et al.  The Redox State of the Cell Regulates the Ligand Binding Affinity of Human Neuroglobin and Cytoglobin* , 2003, Journal of Biological Chemistry.

[43]  Jean-Pierre Timmermans,et al.  A Globin in the Nucleus!* , 2003, Journal of Biological Chemistry.

[44]  J. Paulauskis,et al.  Induction of high mobility group-I(Y) protein by endotoxin and interleukin-1beta in vascular smooth muscle cells. Role in activation of inducible nitric oxide synthase. , 1999, The Journal of biological chemistry.

[45]  Weidong Wang,et al.  NURD, a novel complex with both ATP-dependent chromatin-remodeling and histone deacetylase activities. , 1998, Molecular cell.

[46]  A. Riggs,et al.  Identification of Myoglobin in Human Smooth Muscle* , 1998, The Journal of Biological Chemistry.

[47]  Hong Wang,et al.  Enhancement of Serum-response Factor-dependent Transcription and DNA Binding by the Architectural Transcription Factor HMG-I(Y)* , 1998, The Journal of Biological Chemistry.

[48]  D. Fujiwara,et al.  Leptomycin B targets a regulatory cascade of crm1, a fission yeast nuclear protein, involved in control of higher order chromosome structure and gene expression. , 1994, The Journal of biological chemistry.

[49]  R. Hertzberg,et al.  On the mechanism of topoisomerase I inhibition by camptothecin: evidence for binding to an enzyme-DNA complex. , 1989, Biochemistry.

[50]  D. Conti,et al.  The Hemoglobin Homolog Cytoglobin in Smooth Muscle Inhibits Apoptosis and Regulates Vascular Remodeling , 2017, Arteriosclerosis, thrombosis, and vascular biology.

[51]  K. Wakasa,et al.  Tumorigenesis and Neoplastic Progression Promotion of Liver and Lung Tumorigenesis in DEN-Treated Cytoglobin-Deficient Mice , 2011 .

[52]  M. Štros.  HMGB proteins: interactions with DNA and chromatin. , 2010, Biochimica et biophysica acta.