Human fetal globin gene expression is regulated by LYAR.
Bottom Line: We found that PRMT5 binding on the proximal γ-promoter was LYAR-dependent.We also found that LYAR repressed human fetal globin gene expression in both K562 cells and primary human adult erythroid progenitor cells.Thus, these data indicate that LYAR acts as a novel transcription factor that binds the γ-globin gene, and is essential for silencing the γ-globin gene.
Affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210093, China.Show MeSH
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Mentions: In order to test whether LYAR together with PRMT5 functions to transcriptionally regulate gene activity by binding to GGTTAT, we constructed a luciferase reporter gene driven by DNase hypersensitive site 2 (HS2) of the β-globin locus and a minimal γ-promoter (nucleotide −130 to +40) with either wild-type (WT) or a mutant LYAR-binding site (Figure 3A). When the WT reporter was co-transfected with expression vector containing HA-tagged LYAR or FLAG-tagged PRMT5 or both into 293T cells, a significant decrease in relative luciferase activity was observed compared to the empty vector (Figure 3B). However, no significant change was found when the mutant LYAR-binding site reporter was used (Figure 3C). The expression of the exogenous protein was verified by western blot analysis (Figure 3D). This result demonstrated that LYAR could regulate transcription through binding GGTTAT.
Affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210093, China.