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PLoS By Category | Recent PLoS Articles
Hematology - Molecular Biology

MicroRNA-96 Directly Inhibits ?-Globin Expression in Human Erythropoiesis
Published: Thursday, July 28, 2011
Author: Imane Azzouzi et al.

by Imane Azzouzi, Hansjoerg Moest, Jeannine Winkler, Jean-Claude Fauchère, André P. Gerber, Bernd Wollscheid, Markus Stoffel, Markus Schmugge, Oliver Speer

Fetal hemoglobin, HbF (a2?2), is the main hemoglobin synthesized up to birth, but it subsequently declines and adult hemoglobin, HbA (a2ß2), becomes predominant. Several studies have indicated that expression of the HbF subunit ?-globin might be regulated post-transcriptionally. This could be confered by ~22-nucleotide long microRNAs that associate with argonaute proteins to specifically target ?-globin mRNAs and inhibit protein expression. Indeed, applying immunopurifications, we found that ?-globin mRNA was associated with argonaute 2 isolated from reticulocytes that contain low levels of HbF (<1%), whereas association was significantly lower in reticulocytes with high levels of HbF (90%). Comparing microRNA expression in reticulocytes from cord blood and adult blood, we identified several miRNAs that were preferentially expressed in adults, among them miRNA-96. The overexpression of microRNA-96 in human ex vivo erythropoiesis decreased ?-globin expression by 50%, whereas the knock-down of endogenous microRNA-96 increased ?-globin expression by 20%. Moreover, luciferase reporter assays showed that microRNA-96 negatively regulates expression of ?-globin in HEK293 cells, which depends on a seedless but highly complementary target site located within the coding sequence of ?-globin. Based on these results we conclude that microRNA-96 directly suppresses ?-globin expression and thus contributes to HbF regulation.
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