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J. Biol. Chem., Vol. 278, Issue 49, 48704-48712, December 5, 2003

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DNase I Hypersensitivity Patterns of the Serglycin Proteoglycan Gene in Resting and Phorbol 12-Myristate 13-Acetate-stimulated Human Erythroleukemia (HEL), CHRF 288-11, and HL-60 Cells Compared with Neutrophils and Human Umbilical Vein Endothelial Cells^*

Patria Castronuevo, Michael A. Thornton, Lois E. McCarthy, Joanne Klimas, and Barbara P. Schick¶

From the Cardeza Foundation for Hematologic Research, Department of Medicine, Jefferson Medical College of Thomas Jefferson University and Department of Pediatrics, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19107

We mapped the DNase I-hypersensitive sites (DHSS) of the serglycin gene in resting and phorbol 12-myristate 13-acetate (PMA)-stimulated human erythroleukemia (HEL) and CHRF 288-11 cells, which have megakaryocytic characteristics, and HL-60 promyelocytic leukemia cells. We compared these DHSS with those of normal primary neutrophils and human umbilical vein endothelial cells. Several DHSS appear to be involved in regulating the level of endogenous expression and in the PMA response of hematopoietic cell lines. A DHSS unique to resting HL-60 cells and induced in CHRF 288-11 by PMA may explain the high degree of endogenous expression in HL-60 relative to HEL and CHRF (Schick, B. P., Petrushina, I., Brodbeck, K. C., and Castronuevo, P. (2001) J. Biol. Chem. 276, 24726–24735). A total of 4 DHSS in intron 1 and 6 in intron 2 are associated with the PMA response in a cell-specific manner. A DHSS in the 5'-flanking region and another in intron 1 lie in areas that have high homology with the orthologous murine serglycin locus and are rich in potential transcription factor binding sites. One DHSS in intron 1 and one in intron 2 are located within Alu repeats. Two DHSS found in DNA of normal primary neutrophils were different from those of the cell lines. One DHSS in exon 2 unique to neutrophils correlated with a previously unrecognized alternative splicing that removes exon 2. Human umbilical vein endothelial cells had a DHSS in intron 1 that was common with the cell lines. The different patterns of DHSS exhibited by the cells studied suggest that cell- and differentiation-specific alterations in chromatin structure may control serglycin gene expression.

Received for publication, September 15, 2003

^* This work was supported by United States Public Health Service Grants RO1-HL29282 (to B. P. S.) and T32-HL07821 (to P. C. and L. E. M.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed: Rm. 709 Curtis Bldg., Cardeza Foundation for Hematologic Research, Thomas Jefferson University, 1015 Walnut St., Philadelphia, PA 19107. Tel.: 215-955-6312; Fax: 215-955-2366; E-mail: barbara.schick{at}jefferson.edu.

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