A distal single nucleotide polymorphism alters long-range regulation of the PU.1 gene in acute myeloid leukemia
Ulrich Steidl, … , Frank Griesinger, Daniel G. Tenen
Ulrich Steidl, … , Frank Griesinger, Daniel G. Tenen
Published September 4, 2007
Citation Information: J Clin Invest. 2007;117(9):2611-2620. https://doi.org/10.1172/JCI30525.
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A distal single nucleotide polymorphism alters long-range regulation of the PU.1 gene in acute myeloid leukemia

Abstract

Targeted disruption of a highly conserved distal enhancer reduces expression of the PU.1 transcription factor by 80% and leads to acute myeloid leukemia (AML) with frequent cytogenetic aberrations in mice. Here we identify a SNP within this element in humans that is more frequent in AML with a complex karyotype, leads to decreased enhancer activity, and reduces PU.1 expression in myeloid progenitors in a development-dependent manner. This SNP inhibits binding of the chromatin-remodeling transcriptional regulator special AT-rich sequence binding protein 1 (SATB1). Overexpression of SATB1 increased PU.1 expression, and siRNA inhibition of SATB1 downregulated PU.1 expression. Targeted disruption of the distal enhancer led to a loss of regulation of PU.1 by SATB1. Interestingly, disruption of SATB1 in mice led to a selective decrease of PU.1 RNA in specific progenitor types (granulocyte-macrophage and megakaryocyte-erythrocyte progenitors) and a similar effect was observed in AML samples harboring this SNP. Thus we have identified a SNP within a distal enhancer that is associated with a subtype of leukemia and exerts a deleterious effect through remote transcriptional dysregulation in specific progenitor subtypes.

Authors

Ulrich Steidl, Christian Steidl, Alexander Ebralidze, Björn Chapuy, Hye-Jung Han, Britta Will, Frank Rosenbauer, Annegret Becker, Katharina Wagner, Steffen Koschmieder, Susumu Kobayashi, Daniel B. Costa, Thomas Schulz, Karen B. O’Brien, Roel G.W. Verhaak, Ruud Delwel, Detlef Haase, Lorenz Trümper, Jürgen Krauter, Terumi Kohwi-Shigematsu, Frank Griesinger, Daniel G. Tenen

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Figure 3

The SNP in the first homology region of the URE of PU.1 diminishes binding of SATB1.

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The SNP in the first homology region of the URE of PU.1 diminishes bindi...
(A) Chromatin immunoprecipitation shows SATB1 binding to the URE in U937 and HL60 cells. The genomic region of the putative SATB1 binding site was PCR amplified after reverse crosslink of the immunoprecipitates. An input control and precipitates utilizing a SATB1 antibody, no antibody, or a nonspecific control antibody are shown. PCR products were verified by sequencing. (B) EMSA utilizing nuclear extracts of U937 cells and gel-purified probes (WT probe, 32P, and SNP probe, 32P) covering the SATB1 binding site are shown. The wild-type probe (WT oligo) and a previously described SATB1 binding probe (SATB1 IgH site) were used for competition. A SATB1 antibody was used for supershift. A labeled Sp1 binding probe served as a loading control in 2 lanes. The 32P-labeled SATB1 IgH site served as positive control. The presence (+) or absence (–) of the respective reagents is indicated for each lane. The probes are shown below the gel.