This proposal will investigate the role of a newly rediscovered posttranslational modification-N-terminal protein arginylation-in cardiovascular development. Prior publications by the PI demonstrate that protein arginylation is critical for cardiovascular development and that knockout of the protein arginylation enzyme ATE1 results in embryonic lethality and severe cardiovascular phenotypes in mice that closely resemble the most commonly occurring congenital heart defects in humans. The proposed research is aimed to gain a general understanding of the role of protein arginylation in cardiovascular development and gain insights into possible mechanisms and ways of treatment of the congenital heart diseases, through the following three specific aims: (1) To test the hypothesis that arginylation regulates cell migration and specification and possibly affects the rates of apopotosis during heart development, using live observations of cultured embryonic cells, immunohistochemistry, insitu hybridization, and microarray profiling;(2) To identify cell lineages responsible for the Ate1 -/- developmental phenotype and to test the hypothesis that this phenotype is due at least in part to changes in cells of the neural creast and endothelial lineages and cardiac myocytes, using conditional mouse ATE1 knockouts;and (3) To identify proteins arginylated in vivo in cardiovascular tissues and to define the possible molecular mechanisms of the Ate1-/- cardiovascular phenotype, using biochemical analysis and mass spectrometry. Taken together, the proposed experiments will allow to identify the specific pathways in cardiovascular development controlled by protein arginylation and to gain insights into the molecular mechanisms of this control at the gene expression and protein level. Relevance to public health: Ate1-/- cardiovascular phenotypes resemble commonly occurring human congenital heart diseases, including ventricular and atrial septal defects (VSD and ASD), persistent truncus artheriosis (PTA), and DiGeorge syndrome. The proposed research is aimed to address the previously unexplored mechanisms of these diseases and provide insights into the new possibilities of their treatment.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL084419-05
Application #
8010639
Study Section
Cardiovascular Differentiation and Development Study Section (CDD)
Program Officer
Schramm, Charlene A
Project Start
2007-01-01
Project End
2013-12-31
Budget Start
2011-01-01
Budget End
2013-12-31
Support Year
5
Fiscal Year
2011
Total Cost
$393,750
Indirect Cost
Name
University of Pennsylvania
Department
Veterinary Sciences
Type
Schools of Veterinary Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Terman, Jonathan R; Kashina, Anna (2013) Post-translational modification and regulation of actin. Curr Opin Cell Biol 25:30-8
Shabalina, Svetlana A; Spiridonov, Nikolay A; Kashina, Anna (2013) Sounds of silence: synonymous nucleotides as a key to biological regulation and complexity. Nucleic Acids Res 41:2073-94
Ribeiro, Paula A B; Ribeiro, Jorge P; Minozzo, Fábio C et al. (2013) Contractility of myofibrils from the heart and diaphragm muscles measured with atomic force cantilevers: effects of heart-specific deletion of arginyl-tRNA-protein transferase. Int J Cardiol 168:3564-71
Saha, Sougata; Wang, Junling; Buckley, Brian et al. (2012) Small molecule inhibitors of arginyltransferase regulate arginylation-dependent protein degradation, cell motility, and angiogenesis. Biochem Pharmacol 83:866-73
Zhang, Fangliang; Saha, Sougata; Kashina, Anna (2012) Arginylation-dependent regulation of a proteolytic product of talin is essential for cell-cell adhesion. J Cell Biol 197:819-36
Kurosaka, Satoshi; Leu, N Adrian; Pavlov, Ivan et al. (2012) Arginylation regulates myofibrils to maintain heart function and prevent dilated cardiomyopathy. J Mol Cell Cardiol 53:333-41
Saha, Sougata; Kashina, Anna (2011) Posttranslational arginylation as a global biological regulator. Dev Biol 358:1-8
Wang, Junling; Han, Xuemei; Saha, Sougata et al. (2011) Arginyltransferase is an ATP-independent self-regulating enzyme that forms distinct functional complexes in vivo. Chem Biol 18:121-30
Saha, Sougata; Wong, Catherine C L; Xu, Tao et al. (2011) Arginylation and methylation double up to regulate nuclear proteins and nuclear architecture in vivo. Chem Biol 18:1369-78
Saha, Sougata; Mundia, Maureen M; Zhang, Fangliang et al. (2010) Arginylation regulates intracellular actin polymer level by modulating actin properties and binding of capping and severing proteins. Mol Biol Cell 21:1350-61

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