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A role for Gcn5 in cardiomyocyte differentiation of rat mesenchymal stem cells
A role for Gcn5 in cardiomyocyte differentiation of rat mesenchymal stem cells
- Source :
- Molecular and Cellular Biochemistry. 345:309-316
- Publication Year :
- 2010
- Publisher :
- Springer Science and Business Media LLC, 2010.
-
Abstract
- MSCs possess the capacity of self-renewal and potential of differentiation into various kinds of specialized tissue cells including myocardiocytes. From self-renewing to oriented differentiation, chromatin is remodeled into heritable states that allow activation or maintain the repression of regulatory genes, which means specific genes in self-renewing switched off and specific genes in oriented differentiation activated (Bernstein et al. Cell 125:315-326, 2006). These epigenetic states are established and controlled largely by specific patterns of histone posttranslational modifications, in particular, histone acetylation (Li Nat Rev Genet 3:662-673, 2002). In cardiomyocyte differentiation of rat MSCs, we focused on Gcn5, which linked a known transcriptional coactivator with catalytic histone acetyltransferase activity (Brownell et al. Cell 84:843-851, 1996). To clarify participatory in vivo role of Gcn5, using an RNA interference (RNAi) strategy employing shRNA to specifically knockdown Gcn5 expression in MSCs, we found that HAT activity altered dynamically depended on the inhibition of Gcn5 during MSCs differentiation. Chromatin immunoprecipitation (ChIP) assay showed the increased binding of acetyl histone H3 to the early cardiomyocyte-specific genes GATA4 and NKx2.5 promoters in cardiomyocyte differentiation of MSCs by 5-azacytidine inducing, whereas the decreased binding with lower Gcn5 expression. Cell ultrastructure analysis revealed that MSCs induced by 5-azacytidine possess morphological characteristics of cardiomyocyte cells. The shape of MSCs transfected by Gcn5 RNAi was similar to normal MSCs, but the chromatin showed heavy electron-density and a hard-packed structure. This intermediate state of chromatin may be an inactive part of MSCs differentiation. These results demonstrate that Gcn5, possessing acetyltransferase activity, is involved in regulating chromatin configuration around GATA4 and NKx2.5 in cardiomyocyte differentiation of rat MSCs by changing the level of histone acetylation. HAT activity depending on Gcn5 is important in differentiation of MSCs into cardiomyocytes as a consequence of the remodeling of the chromatin configuration caused by modification of histone H3.
- Subjects :
- Male
Clinical Biochemistry
Chromatin remodeling
Histones
Histone H3
Histone H2A
Animals
Humans
Histone acetyltransferase activity
Myocytes, Cardiac
Epigenetics
RNA, Small Interfering
Rats, Wistar
Molecular Biology
Histone Acetyltransferases
biology
Acetylation
Cell Differentiation
Mesenchymal Stem Cells
Cell Biology
General Medicine
Molecular biology
Rats
Chromatin
Cell biology
enzymes and coenzymes (carbohydrates)
Histone
biology.protein
Chromatin immunoprecipitation
Subjects
Details
- ISSN :
- 15734919 and 03008177
- Volume :
- 345
- Database :
- OpenAIRE
- Journal :
- Molecular and Cellular Biochemistry
- Accession number :
- edsair.doi.dedup.....83571429899e1bd4c238f6333eaf3d55
- Full Text :
- https://doi.org/10.1007/s11010-010-0586-3