Defective sumoylation pathway directs congenital heart disease.

Congenital heart defects (CHDs) are the most common of all birth defects, yet molecular mechanism(s) underlying highly prevalent atrial septal defects (ASDs) and ventricular septal defects (VSDs) have remained elusive. We demonstrate the indispensability of "balanced" posttranslational small ubiquitin-like modifier (SUMO) conjugation-deconjugation pathway for normal cardiac development. Both hetero- and homozygous SUMO-1 knockout mice exhibited ASDs and VSDs with high mortality rates, which were rescued by cardiac reexpression of the SUMO-1 transgene. Because SUMO-1 was also involved in cleft lip/palate in human patients, the previous findings provided a powerful rationale to question whether SUMO-1 was mutated in infants born with cleft palates and ASDs. Sequence analysis of DNA from newborn screening blood spots revealed a single 16 bp substitution in the SUMO-1 regulatory promoter of a patient displaying both oral-facial clefts and ASDs. Diminished sumoylation activity whether by genetics, environmental toxins, and/or pharmaceuticals may significantly contribute to susceptibility to the induction of congenital heart disease worldwide. Birth Defects Research (Part A) 2011. © 2011 Wiley-Liss, Inc.

[1]  M. Dasso,et al.  Modification in reverse: the SUMO proteases. , 2007, Trends in biochemical sciences.

[2]  E. Miska,et al.  The SUMO E3 ligase RanBP2 promotes modification of the HDAC4 deacetylase , 2002, The EMBO journal.

[3]  M. Dasso,et al.  Distinct in vivo dynamics of vertebrate SUMO paralogues. , 2004, Molecular biology of the cell.

[4]  M. Shirakawa,et al.  Small ubiquitin-like modifier 1 (SUMO-1) modification of the synergy control motif of Ad4 binding protein/steroidogenic factor 1 (Ad4BP/SF-1) regulates synergistic transcription between Ad4BP/SF-1 and Sox9. , 2004, Molecular endocrinology.

[5]  R. Schwartz,et al.  Rho kinases play an obligatory role in vertebrate embryonic organogenesis. , 2001, Development.

[6]  T. Pawełczyk,et al.  Expression level of Ubc9 protein in rat tissues. , 2003, Acta Biochimica Polonica.

[7]  S. Izumo,et al.  The cardiac homeobox gene Csx/Nkx2.5 lies genetically upstream of multiple genes essential for heart development. , 1999, Development.

[8]  Xuedong Liu,et al.  Ubc9 expression is essential for myotube formation in C2C12. , 2006, Experimental cell research.

[9]  A. Karsan,et al.  Notch Signaling in Cardiac Development , 2008, Circulation research.

[10]  R. Schwartz,et al.  Regulation of Cardiac Specific nkx2.5 Gene Activity by Small Ubiquitin-like Modifier* , 2008, Journal of Biological Chemistry.

[11]  A. Baldini.  Dissecting contiguous gene defects: TBX1. , 2005, Current opinion in genetics & development.

[12]  J. Molkentin,et al.  The transcription factors GATA 4 and GATA 6 regulate cardiomyocyte hypertrophy in vitro and in vivo , 2001 .

[13]  S. Bhattacharya,et al.  Genetic Mechanisms Controlling Cardiovascular Development , 2008, Annals of the New York Academy of Sciences.

[14]  T. Karpova,et al.  Crm1 is a mitotic effector of Ran-GTP in somatic cells , 2005, Nature Cell Biology.

[15]  Joseph C. Wu,et al.  Common congenital heart disorders in adults. , 2004, Current problems in cardiology.

[16]  Benjamin J Bondow,et al.  Loss of both GATA4 and GATA6 blocks cardiac myocyte differentiation and results in acardia in mice. , 2008, Developmental biology.

[17]  H. Saitoh,et al.  Functional Heterogeneity of Small Ubiquitin-related Protein Modifiers SUMO-1 versus SUMO-2/3* , 2000, The Journal of Biological Chemistry.

[18]  R. Schwartz,et al.  SUMO-specific protease 2 is essential for suppression of polycomb group protein-mediated gene silencing during embryonic development. , 2010, Molecular cell.

[19]  W. Claycomb,et al.  Cardiac physiology at the cellular level: use of cultured HL-1 cardiomyocytes for studies of cardiac muscle cell structure and function. , 2004, American journal of physiology. Heart and circulatory physiology.

[20]  E. Yeh,et al.  SUMOylation and De-SUMOylation: Wrestling with Life's Processes* , 2009, Journal of Biological Chemistry.

[21]  K. Shuai,et al.  Resolution of Sister Centromeres Requires RanBP2-Mediated SUMOylation of Topoisomerase IIα , 2008, Cell.

[22]  J. Hoffman,et al.  The incidence of congenital heart disease. , 2002, Journal of the American College of Cardiology.

[23]  J. McNally,et al.  Modification of de novo DNA methyltransferase 3a (Dnmt3a) by SUMO-1 modulates its interaction with histone deacetylases (HDACs) and its capacity to repress transcription. , 2004, Nucleic acids research.

[24]  Pier Paolo Pandolfi,et al.  The SUMO pathway is essential for nuclear integrity and chromosome segregation in mice. , 2005, Developmental cell.

[25]  W. R. MacLellan,et al.  Cardiac myocyte cell cycle control in development, disease, and regeneration. , 2007, Physiological reviews.

[26]  J. Schmitt,et al.  A Murine Model of Holt-Oram Syndrome Defines Roles of the T-Box Transcription Factor Tbx5 in Cardiogenesis and Disease , 2001, Cell.

[27]  M. Dasso.  Emerging roles of the SUMO pathway in mitosis , 2008, Cell Division.

[28]  M. Mann,et al.  Distinct and Overlapping Sets of SUMO-1 and SUMO-2 Target Proteins Revealed by Quantitative Proteomics*S , 2006, Molecular & Cellular Proteomics.

[29]  D. Srivastava,et al.  Serum response factor orchestrates nascent sarcomerogenesis and silences the biomineralization gene program in the heart , 2008, Proceedings of the National Academy of Sciences.

[30]  M. Nakao,et al.  Serum response factor is modulated by the SUMO-1 conjugation system. , 2003, Biochemical and biophysical research communications.

[31]  K. Helin,et al.  The Polycomb Repressive Complex 2 Is a Potential Target of SUMO Modifications , 2008, PloS one.

[32]  M. Matunis,et al.  SUMO-2/3 modification and binding regulate the association of CENP-E with kinetochores and progression through mitosis. , 2008, Molecules and Cells.

[33]  J. Toppari,et al.  Sumo-1 Function Is Dispensable in Normal Mouse Development , 2008, Molecular and Cellular Biology.

[34]  B. Spiegelman,et al.  C/EBPβ Controls Exercise-Induced Cardiac Growth and Protects against Pathological Cardiac Remodeling , 2010, Cell.

[35]  S. Lockett,et al.  Loss of SUMO1 in mice affects RanGAP1 localization and formation of PML nuclear bodies, but is not lethal as it can be compensated by SUMO2 or SUMO3 , 2008, Journal of Cell Science.

[36]  Jennifer J. Lund,et al.  SUMO1 Haploinsufficiency Leads to Cleft Lip and Palate , 2006, Science.

[37]  C. Deng,et al.  Disruption of Smad4 in neural crest cells leads to mid-gestation death with pharyngeal arch, craniofacial and cardiac defects. , 2008, Developmental biology.

[38]  R. Schwartz,et al.  Myocardin Sumoylation Transactivates Cardiogenic Genes in Pluripotent 10T1/2 Fibroblasts , 2006, Molecular and Cellular Biology.

[39]  N J Izzo,et al.  HL-1 cells: a cardiac muscle cell line that contracts and retains phenotypic characteristics of the adult cardiomyocyte. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[40]  Jun Wang.  SUMO conjugation and cardiovascular development. , 2009, Frontiers in bioscience.

[41]  E. Yeh,et al.  Ubiquitin-like proteins: new wines in new bottles. , 2000, Gene.

[42]  Jun Wang,et al.  SUMO-1 Modification Activated GATA4-dependent Cardiogenic Gene Activity* , 2004, Journal of Biological Chemistry.

[43]  E. Stanley,et al.  Cardiac Septal and Valvular Dysmorphogenesis in Mice Heterozygous for Mutations in the Homeobox Gene Nkx2-5 , 2000, Circulation research.

[44]  R. Barlow,et al.  Heterogeneity of Genetic Modifiers Ensures Normal Cardiac Development , 2010, Circulation.

[45]  Jingde Zhu,et al.  Stabilization of PML nuclear localization by conjugation and oligomerization of SUMO-3 , 2005, Oncogene.

[46]  Jian Li,et al.  Myocardin is required for cardiomyocyte survival and maintenance of heart function , 2009, Proceedings of the National Academy of Sciences.

[47]  R. Schwartz,et al.  Sumoylation and regulation of cardiac gene expression. , 2010, Circulation research.

[48]  R. DePinho,et al.  SUMO-1 Modification of Histone Deacetylase 1 (HDAC1) Modulates Its Biological Activities* , 2002, The Journal of Biological Chemistry.

[49]  Y. Nakamura,et al.  Cloning, expression, and mapping of UBE2I, a novel gene encoding a human homologue of yeast ubiquitin-conjugating enzymes which are critical for regulating the cell cycle. , 1996, Cytogenetics and cell genetics.

[50]  R. Schwartz,et al.  Cardiac-Specific Deletion of Gata4 Reveals Its Requirement for Hypertrophy, Compensation, and Myocyte Viability , 2006, Circulation research.

[51]  M. Tatham,et al.  Unique binding interactions among Ubc9, SUMO and RanBP2 reveal a mechanism for SUMO paralog selection , 2005, Nature Structural &Molecular Biology.

[52]  Jonathan C. Cohen,et al.  GATA4 mutations cause human congenital heart defects and reveal an interaction with TBX5 , 2003, Nature.

[53]  R. Schwartz,et al.  Embryonic expression of an Nkx2‐5/Cre gene using ROSA26 reporter mice , 2001, Genesis.

[54]  M. Kagey,et al.  The Polycomb Protein Pc2 Is a SUMO E3 , 2003, Cell.

[55]  Erica S. Johnson,et al.  Protein modification by SUMO. , 2004, Annual review of biochemistry.

[56]  David H Russell,et al.  A Universal Strategy for Proteomic Studies of SUMO and Other Ubiquitin-like Modifiers*S , 2005, Molecular & Cellular Proteomics.

[57]  A. Woollard,et al.  The T-box factor TBX-2 and the SUMO conjugating enzyme UBC-9 are required for ABa-derived pharyngeal muscle in C. elegans. , 2006, Developmental biology.

[58]  Matthias Mann,et al.  A Proteomic Study of SUMO-2 Target Proteins* , 2004, Journal of Biological Chemistry.