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J. Biol. Chem., Vol. 282, Issue 30, 21924-21933, July 27, 2007

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CCAAT Enhancer-binding Protein Regulates Constitutive Gene Expression during Late Stages of Monocyte to Macrophage Differentiation^*

From the Department of Hematology and Oncology, University Hospital Regensburg, 93042 Regensburg, Germany

Human monocyte to macrophage differentiation is accompanied by pronounced phenotypical changes and generally proceeds in the absence of proliferation. The molecular events governing this process are poorly understood. Here, we studied the regulation of the macrophage-specific chitotriosidase (CHIT1) gene promoter to gain insights into the mechanisms of transcriptional control during the differentiation of human blood monocytes into macrophages. We used transient transfections to define a cell type-specific minimal promoter that was mainly dependent on a proximal C/EBP motif that bound multiple C/EBP factors in gel shift assays. In depth analysis of occupied promoter elements using in vivo footprinting and chromatin immunoprecipitation analyses demonstrated the differentiation-associated recruitment of C/EBP and PU.1 at the proximal promoter in parallel with CHIT1 mRNA induction. Notably, the induction of C/EBP promoter binding strongly correlated with increased nuclear levels of Thr-235-phosphorylated C/EBP protein during the differentiation process, whereas C/EBP mRNA and total protein expression remained relatively stable. Our data suggest an important constitutive gene regulatory function for C/EBP in differentiated macrophages but not in human blood monocytes.

Received for publication, December 19, 2006 , and in revised form, May 11, 2007.

^* This work was supported by Deutsche Forschungsgemeinschaft Grant Kr1373/5-3 (to S. W. K. and M. R.). 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.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Figs. 1 and 2 and Table 1.

¹ To whom correspondence should be addressed. Tel.: 49-941-944-5587; Fax: 49-941-944-5593; E-mail: michael.rehli{at}klinik.uni-regensburg.de.

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