Nucleic Acids Research, 1994, Vol. 22, No. 25 5672-5678
© 1994
Articles |
Identification of a DNA binding site for the nuclear factor YY1 in the human GM-CSF core promoter
Laboratory of Experimental Immunology, Biological Response Modifier Program DCT, NCI - FCRDC, Frederick, MD 21702-1201, USA
*To whom correspondence should be addressed
Received August 19, 1994. Revised November 12, 1994. Accepted November 12, 1994.
It has been well documented that the repeated CATT(A/T) sequence, localized between - 64 and - 35 in the human GM-CSF promoter, Is required for the promoter activity, and this region likely serves as a core recognition sequence for a cellular transcription factor. However, the transcription factor that interacts with this site was not Identified. Here, we report that this element contains a binding site for the nuclear factor YY1, which has not been reported to play a role in the regulation of cytokine gene transcription. Results from transient transfection assays of the Jurkat T cell line revealed that this repeated CATT(A/T) element exhibited enhancer activity when linked to both the human IFN
promoter and the TK promoter. Mutation of the YY1 binding site eliminated about 60% of the enhancer activity of the element. We have found that the YY1 binding site could form two specific DNA-protein complexes, A and B, with Jurkat nuclear proteins in the electrophoretic mobility shift assay and that the binding of these complexes correlates with the enhancer activity. UV cross-linking analysis revealed that the A complex is a multl-proteln complex and in addition to YY1, other proteins are required for formation of the protein complex. Cotransfectlon assays with a YY1 expression vector revealed that overexpresslon of YY1 resulted In an inhibitory effect on the repeated CATT(A/T) element, indicating that In addition to YY1, cofactors also are required for the activator function of the A complex.
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