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ShRNA profiling of mouse erythroleukemia (MEL) cells and their corresponding G9a knock downs reveals a dual role for the methyltransferase G9a in the maintenance of beta-globin gene expression

ABSTRACT: MEL clone 745 cells that are blocked at the pro-erythroblast stage serve as a model for terminal erythroid differentiation when treated with DMSO. A stable MEL cell line expressing (Dox)-dependent shRNA sequence targeting different regions of G9a mRNA was established and screened as previously described in (Demers, 2007 PMID17707229). Knock down was induced by adding 5 micrograms/ml final of Dox in the culture medium. For a total of six samples, 10 micro grams of RNA was extracted from each three biological replicates of mouse erythroleukemia (MEL) cells and their corresponding G9a knock downs. The RNA was extracted and purified using the RNeasy Mini Kit (Qiagen) including the on-column DNAse I digestion step. Experiment Overall Design: Mouse erythroleukemia cells: wild type vs G9 knock down. For each microarray hybridization, data analysis was performed using MAS and GCRMA.

ORGANISM(S): Mus musculus

SUBMITTER: OGIC Info Ontario Genomics Innovation Centre (OGIC) 

PROVIDER: E-GEOD-15620 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Dual role for the methyltransferase G9a in the maintenance of beta-globin gene transcription in adult erythroid cells.

Chaturvedi Chandra-Prakash CP   Hosey Alison M AM   Palii Carmen C   Perez-Iratxeta Carolina C   Nakatani Yoshihiro Y   Ranish Jeffrey A JA   Dilworth F Jeffrey FJ   Brand Marjorie M  

Proceedings of the National Academy of Sciences of the United States of America 20091012 43

Using a proteomics screen, we have identified the methyltransferase G9a as an interacting partner of the hematopoietic activator NF-E2. We show that G9a is recruited to the beta-globin locus in a NF-E2-dependent manner and spreads over the entire locus. While G9a is often regarded as a corepressor, knocking down this protein in differentiating adult erythroid cells leads to repression of the adult beta(maj) globin gene and aberrant reactivation of the embryonic beta-like globin gene E(y). While  ...[more]

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