Unknown,Transcriptomics,Genomics,Proteomics

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Analysis of expression levels of genes on chromosome 19 in U937 cells expressing AML1/ETO


ABSTRACT: Approximately 20% of Acute Myelogenous Leukemia (AML) cases carry the t(8;21) translocation, which involves the AML1 and ETO genes, and express the resulting AML1/ETO fusion protein that functions as a transcriptional repressor by recruiting NCoR/SMRT/HDAC complexes to DNA. We used ChIP-chip to identify the determinants of AML1/ETO binding on a contiguous DNA region (chromosome 19). In order to correlate transcription factor binding to gene expression, we evaluated expression levels of genes localized on chromosome 19 by expression tiling. Keywords: Gene expression A U937 cell line that conditionally expresses HA-tagged AML1/ETO under the control of the mouse metallothionine promoter (U937-A1E) (Alcalay et al., J.Clin.Invest, 2003,112, 1751-1761) was used. A cell line carrying the empty vector was used as control. Cell lines were treated for 8h with 100uM ZnSO4 to induce transgene expression in U937-A1E. For each of the U937 cell lines (AML1/ETO and Mt), three independent RNA extractions were performed, and an equal quantity of each of the three RNA preparations was then mixed to generate an RNA pool for each sample.

ORGANISM(S): Homo sapiens

SUBMITTER: Myriam Alcalay 

PROVIDER: E-GEOD-10579 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

AML1/ETO oncoprotein is directed to AML1 binding regions and co-localizes with AML1 and HEB on its targets.

Gardini Alessandro A   Cesaroni Matteo M   Luzi Lucilla L   Okumura Akiko J AJ   Biggs Joseph R JR   Minardi Simone P SP   Venturini Elisa E   Zhang Dong-Er DE   Pelicci Pier Giuseppe PG   Alcalay Myriam M  

PLoS genetics 20081128 11


A reciprocal translocation involving chromosomes 8 and 21 generates the AML1/ETO oncogenic transcription factor that initiates acute myeloid leukemia by recruiting co-repressor complexes to DNA. AML1/ETO interferes with the function of its wild-type counterpart, AML1, by directly targeting AML1 binding sites. However, transcriptional regulation determined by AML1/ETO probably relies on a more complex network, since the fusion protein has been shown to interact with a number of other transcriptio  ...[more]

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