Transcriptional regulators Myb and BCL11A interplay with DNA methyltransferase 1 in developmental silencing of embryonic and fetal β-like globin genes.
- Author(s)
- Roosjen, M; McColl, B; Kao, B; Gearing, LJ; Blewitt, ME; Vadolas, J;
- Details
- Publication Year 2013-12-26,Volume 28,Issue #4,Page 1610-20
- Journal Title
- FASEB Journal
- Publication Type
- JOURNAL ARTICLE
- Abstract
- The clinical symptoms of hemoglobin disorders such as β-thalassemia and sickle cell anemia are significantly ameliorated by the persistent expression of γ-globin after birth. This knowledge has driven the discovery of important regulators that silence γ-globin postnatally. Improved understanding of the γ- to β-globin switching mechanism holds the key to devising targeted therapies for β-hemoglobinopathies. To further investigate this mechanism, we used the murine erythroleukemic (MEL) cell line containing an intact 183-kb human β-globin locus, in which the (G)γ- and β-globin genes are replaced by DsRed and eGFP fluorescent reporters, respectively. Following RNA interference (RNAi)-mediated knockdown of two key transcriptional regulators, Myb and BCL11A, we observed a derepression of γ-globin, measured by DsRed fluorescence and qRT-PCR (P<0.001). Interestingly, double knockdown of Myb and DNA methyltransferase 1 (DNMT1) resulted in a robust induction of ε-globin, (up to 20% of total β-like globin species) compared to single knockdowns (P<0.001). Conversely, double knockdowns of BCL11A and DNMT1 enhanced γ-globin expression (up to 90% of total β-like globin species) compared to single knockdowns (P<0.001). Moreover, following RNAi treatment, expression of human β-like globin genes mirrored the expression levels of their endogenous murine counterparts. These results demonstrate that Myb and BCL11A cooperate with DNMT1 to achieve developmental repression of embryonic and fetal β-like globin genes in the adult erythroid environment.-Roosjen, M., McColl, B., Kao, B., Gearing, L. J., Blewitt, M. E., Vadolas, J. Transcriptional regulators Myb and BCL11A interplay with DNA methyltransferase 1 in developmental silencing of embryonic and fetal β-like globin genes.
- Publisher
- FASEB
- Keywords
- RNA interference, epigenetic repressor, molecular mechanism, β-hemoglobinopathies
- Research Division(s)
- Molecular Medicine
- PubMed ID
- 24371119
- Publisher's Version
- https://doi.org/10.1096/fj.13-242669
- Terms of Use/Rights Notice
- Copyright © 2014 by the Federation of American Societies for Experimental Biology
Creation Date: 2014-02-19 11:35:36
Last Modified: 2015-09-04 11:28:40