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J Biol Chem, Vol. 274, Issue 29, 20098-20102, July 16, 1999

Myeloid ELF-1-like Factor Up-regulates Lysozyme Transcription in Epithelial Cells

Hirofumi Kai, Akinori Hisatsune, Takahiro Chihara, Ayako Uto, Ayako Kokusho, Takeshi Miyata, and Carol Basbaum^¶

From the  Department of Pharmacological Sciences, Faculty of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Kumamoto 862, Japan and the ^¶ Department of Anatomy and the Cardiovascular Research Institute, University of California, San Francisco, California 94143

Lysozyme is an important component of innate immunity against common pathogens at mucosal surfaces. We previously cloned and characterized the bovine lysozyme 5A (lys5A) promoter with the purpose of determining cis- and trans-acting elements controlling airway epithelial cell-specific expression. We found that such expression is controlled by protein binding to an ETS consensus sequence located approximately at 46 to 40 bp from the transcription start site. The identity of the ETS-related protein responsible for gene transactivation was unknown. In this study, we screened six ETS-related proteins by transient transfection into epithelial cells and fibroblasts. Results showed that among these factors, the myeloid Elf-1-like factor (MEF) was the most potent. Gel shift analysis of epithelial cell nuclear extracts using a lys5A probe including the ETS-binding site (50/31) yielded a single band with retarded mobility. This band was supershifted by an antibody directed against MEF. Supporting the possibility that MEF is responsible for functional transactivation of lysozyme in epithelial cells, we found that antisense MEF mRNA decreased lys5A promoter activity and that MEF overexpression in stably transfected cells increased lysozyme mRNA and protein expression. We conclude that MEF is required for epithelial cell transactivation of lysozyme.

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