Homeobox Protein HB9 Binds to the Prostaglandin E Receptor 2 Promoter and Inhibits Intracellular cAMP Mobilization in Leukemic Cells

• Published in: The Journal of biological chemistry 2012-11, Vol.287 (48), p.40703-40712
• Main Authors: Wildenhain, Sarah, Ingenhag, Deborah, Ruckert, Christian, Degistirici, Özer, Dugas, Martin, Meisel, Roland, Hauer, Julia, Borkhardt, Arndt
• Format: Article
• Language: English
• Subjects: Cell Line, Tumor; Cyclic AMP - metabolism; Down-Regulation; Gene Expression Regulation, Neoplastic; Gene Regulation; Homeobox; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; Humans; Leukemia; Leukemia, Myeloid, Acute - genetics; Leukemia, Myeloid, Acute - metabolism; Oncogene; Promoter Regions, Genetic; Promoters; Protein Binding; Receptors, Prostaglandin E, EP2 Subtype - genetics; Receptors, Prostaglandin E, EP2 Subtype - metabolism; Transcription Factors; Transcription Factors - genetics; Transcription Factors - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M111.308965

The transcription factor HB9, encoded by the homeobox gene B9 (HLXB9), is involved in the development of pancreatic beta- and motor neuronal cells. In addition, HLXB9 is recurrently rearranged in young children with acute myeloid leukemia characterized by a chromosomal translocation t(7;12)-HLXB9/TEL and concomitant high expression of the unrearranged, wild-type HLXB9 allele. However, target genes of HB9 in hematopoietic cells are not known to date. In this study, we used ChIP-on-chip analysis together with expression profiling and identified PTGER2 (prostaglandin Ereceptor 2) as a target gene of HB9 in a hematopoietic cell line. The functional HB9 homeodomain as well as the HB9 binding domain within the PTGER2 promoter are essential for binding of HB9 to the PTGER2 promoter region and down-regulation of PTGER2 expression. Functionally, HB9 conducted down-regulation of PTGER2 results in a reduced content of intracellular cAMP mobilization and furthermore the decreased PTGER2 gene expression is valid in bone marrow cells from translocation t(7;12) positive patients. Among the primary and secondary target genes of HB9 in the myeloid cell line HL60, 78% of significantly regulated genes are down-regulated, indicating an overall repressive function of HB9. Differentially regulated genes were preferentially confined to pathways involved in cell-adhesion and cell-cell interactions, similar to the gene expression footprint of HLXB9-expressing cells from t(7;12) positive patients. Background: HB9 is highly expressed in translocation t(7;12) positive infant AML. Results: HB9 binds to the PTGER2 promoter, down-regulates PTGER2 gene expression and subsequently represses cAMP synthesis in hematopoietic cells. Conclusion: Expression of HLXB9 represses PTGER2 mediated signaling. Significance: First molecular report of HB9-dependent target gene regulation in hematopoietic cells.