Functional GATA-3 binding sites within murine CD8 alpha upstream regulatory sequences

doi:10.1084/jem.178.3.941

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Functional GATA-3 binding sites within murine CD8 alpha upstream regulatory sequences

DB Landry, JD Engel and R Sen
Rosenstiel Basic Sciences Center, Brandeis University, Waltham, Massachusetts 02254.

Genes encoding the accessory molecules CD8 and CD4 are activated early in thymocyte development, generating CD4+8+ double positive intermediates, which give rise to two functionally distinct mature T cell subsets that express either CD4 or CD8. The mechanisms that govern the activation or suppression of the CD8 gene are likely to be central to the T cell development program. To identify the key regulatory factors, we have initiated an analysis of the transcriptional regulation of the murine CD8 alpha gene. We have identified three CD8+ cell-specific DNAase I hypersensitive sites (HSS) located upstream of the murine CD8 alpha gene. In vitro mobility shift analysis of the -4.0- kb HSS region has revealed multiple binding sites for the T cell- restricted transcription factor GATA-3. In vitro translated murine GATA- 3 binds specifically to both CD8 GATA sites, and coexpression of this factor in transient transfection assays transactivates a reporter construct containing these sequences. These results provide the first evidence for the role of a T cell-restricted factor in the regulation of either CD8 or CD4 genes.
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