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AML1/ETO binding pattern on human promoters

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 microarrays to identify human promoters bound by AML1/ETO in U937 cells. Keywords: ChIP-chip 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. Cells were treated for 8h with 100uM ZnSO4 to induce transgene expression, and ChIP was performed using an anti-HA antibody. ChIP products were then PCR amplified, labeled with Cy3/Cy5 fluorescent dyes and hybridized to the NimbleGen HG17 Human Promoter 2 Array set, which explores 4 kb upstream and 1 kb downstream the transcription start site (TSS) of 24,434 annotated genes. Two biological replicates were prepared and hybridized to independent array sets. U937-Mt cells, which carry the empty vector, served as control for non-specific binding of the anti-HA antibody.

ORGANISM(S): Homo sapiens

SUBMITTER: Myriam Alcalay 

PROVIDER: E-GEOD-10531 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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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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