Functional characterization of the CCL25 promoter in small intestinal epithelial cells suggests a regulatory role for caudal-related homeobox (Cdx) transcription factors.
The chemokine CCL25 is selectively and constitutively expressed in the small intestinal epithelium and plays an important role in mediating lymphocyte recruitment to this site. In this study, we demonstrate that CCL25 expression in murine small intestinal epithelial cells is independent of signaling through the lymphotoxin beta receptor and is not enhanced by inflammatory stimuli, pathways involved in driving the expression of most other chemokines. We define a transcriptional start site in the CCL25 gene and a region -141 to -5 proximal of exon 1 that is required for minimal promoter activity in the small intestinal epithelial cell lines, MODE-K and mICc12. These cell lines expressed far less CCL25 mRNA than freshly isolated small intestinal epithelial cells indicating that they are missing important factors driving CCL25 expression. The CCL25 promoter contained putative binding sites for the intestinal epithelial-associated Caudal-related homeobox (Cdx) transcription factors Cdx-1 and Cdx-2, and small intestinal epithelial cells but not MODE-K and mICc12 cells expressed Cdx-1 and Cdx-2. EMSA analysis demonstrated that Cdx proteins were present in nuclear extracts from freshly isolated small intestinal epithelial cells but not in MODE-K or mICcl2 cells, and bound to putative Cdx sites within the CCL25 promoter. Finally, cotransfection of MODE-K cells with Cdx transcription factors significantly increased CCL25 promoter activity as well as endogenous CCL25 mRNA levels. Together these results demonstrate a unique pattern of regulation for CCL25 and suggest a role for Cdx proteins in regulating CCL25 transcription.
Ericsson
Anna
A
Immunology Section, Stem Cell Center, Biomedical Centre I-13, Lund University, S-22184 Lund, Sweden.
Kotarsky
Knut
K
Svensson
Marcus
M
Sigvardsson
Mikael
M
Agace
William
W
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D000071616:CDX2 Transcription Factor; D002478:Cells, Cultured; D019742:Chemokines, CC / Q000235:genetics*; D017124:Conserved Sequence; D004847:Epithelial Cells / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000235:genetics*; D018398:Homeodomain Proteins / Q000378:metabolism*; D006706:Homeostasis; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D007421:Intestine, Small / Q000166:cytology*; D008214:Lymphocytes / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics; D020218:Response Elements; D014157:Transcription Factors / Q000378:metabolism*
C108074:CCL25 protein, human; D000071616:CDX2 Transcription Factor; C491541:Ccl25 protein, mouse; C491272:Cdx1 protein, mouse; C487514:Cdx2 protein, mouse; D019742:Chemokines, CC; D018398:Homeodomain Proteins; D018836:Inflammation Mediators; D012333:RNA, Messenger; D014157:Transcription Factors