Abstract

Activation of smooth muscle cell (SMC) proliferation in response to vascular injury is a widely appreciated vascular problem in clinical medicine, but its basis remains obscure. Our hypothesis is that abnormal posttranslational modification of proteins makes a major contribution to the initiation of activated smooth muscle cellular proliferation associated with vascular neointimal formation following vascular injury. We are particularly interested in the possibility that O-GlcNAcylation of YY1, significantly elevated in the neontimal smooth muscle cells, regulates the YY1-Rb interaction and downstream transcription promoting gene expression that triggers smooth muscle cellular proliferation. We are interested in the possibility that the phosphorylation of Rb at specific amino acids also inhibits the YY1-Rb complexation contributing to the activation of smooth muscle cellular proliferation. The goal of this project is to identify how O-glycosylation of YY1 regulates the YY1-Rb interaction and the transcriptional changes that occur in SMC transition from quiescence to active proliferation.
Aim 1 : Determine the specific sites of YY1 O-GlcNAcylation in growth arrested and in growth stimulated SMC cultures. The position of O-GlcNAc attachments to YY1 in the fractions will be determined by applying a combination of automated triple quadruple and ion trapping tandem mass spectroscopy.
Aim 2 : Analysis of YY1 O-GlcNAcylation in response to vascular balloon injury in a rat carotid artery model. The rate and the position of YY1 O-glycosylation 2 days, 4 days, and 8 days after rat balloon injury will be determined.
Aim 3 : Structure-based site-directed mutagenesis for the identification of the YY1-Rb binding surface and its regulation by Rb phosphorylation. The phosphorylated Ser/Thr residues on pRb that inhibit YY1 binding will be identified.
Aim 4 : Determine how O-GlcNAcylation influences cellular YY1 functions applying flow cytometry, gene expression regulation, and chromatin immunoprecipitation (ChIP).

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
2R01HL062458-05A2
Application #
6970316
Study Section
Vascular Cell and Molecular Biology Study Section (VCMB)
Program Officer
Rabadan-Diehl, Cristina
Project Start
2000-02-01
Project End
2009-01-31
Budget Start
2005-09-01
Budget End
2006-01-31
Support Year
5
Fiscal Year
2005
Total Cost
$390,000
Indirect Cost

  Institution

Name
Beth Israel Deaconess Medical Center
Department
Type
DUNS #
071723621
City
Boston
State
MA
Country
United States
Zip Code
02215

  Publications

Wilson, Jamie L; Rupasinghe, Chamila; Usheva, Anny et al. (2015) Modulating the dysregulated migration of pulmonary arterial hypertensive smooth muscle cells with motif mimicking cell permeable peptides. Curr Top Pept Protein Res 16:1-17
Nowak-Machen, Martina; Lange, Martin; Exley, Mark et al. (2015) Lysophosphatidic acid generation by pulmonary NKT cell ENPP-2/autotaxin exacerbates hyperoxic lung injury. Purinergic Signal 11:455-61
Duan, Jubao; Shi, Jianxin; Fiorentino, Alessia et al. (2014) A rare functional noncoding variant at the GWAS-implicated MIR137/MIR2682 locus might confer risk to schizophrenia and bipolar disorder. Am J Hum Genet 95:744-53
Bai, Aiping; Moss, Alan; Kokkotou, Efi et al. (2014) CD39 and CD161 modulate Th17 responses in Crohn's disease. J Immunol 193:3366-77
Gelev, Vladimir; Zabolotny, Janice M; Lange, Martin et al. (2014) A new paradigm for transcription factor TFIIB functionality. Sci Rep 4:3664
Lange, Martin; Fujikawa, Tatsuya; Koulova, Anna et al. (2014) Arterial territory-specific phosphorylated retinoblastoma protein species and CDK2 promote differences in the vascular smooth muscle cell response to mitogens. Cell Cycle 13:315-23
Sun, Xiaofeng; Han, Lihui; Seth, Pankaj et al. (2013) Disordered purinergic signaling and abnormal cellular metabolism are associated with development of liver cancer in Cd39/ENTPD1 null mice. Hepatology 57:205-16
Nowak-Lovato, Kristy; Alexandrov, Ludmil B; Banisadr, Afsheen et al. (2013) Binding of nucleoid-associated protein fis to DNA is regulated by DNA breathing dynamics. PLoS Comput Biol 9:e1002881
Alexandrov, Boian S; Phipps, M Lisa; Alexandrov, Ludmil B et al. (2013) Specificity and heterogeneity of terahertz radiation effect on gene expression in mouse mesenchymal stem cells. Sci Rep 3:1184
Pettengill, Matthew; Robson, Simon; Tresenriter, Megan et al. (2013) Soluble ecto-5'-nucleotidase (5'-NT), alkaline phosphatase, and adenosine deaminase (ADA1) activities in neonatal blood favor elevated extracellular adenosine. J Biol Chem 288:27315-26

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