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Nucleic Acids Research, 1994, Vol. 22, No. 3 457-464
© 1994

MOLECULAR BIOLOGY

An S1 nuclease-sensitive region in the first intron of human platelet-derived growth factor A-chain gene contains a negatively acting cell type-specific regulatory element

Zhao-Yi Wang1, Nobuyoshi Masaharu1, Qing-Qing Qiu1, Yasuo Takimato1,+ and Thomas F. Deuel1,2,*

1Department of Medicine, The Jewish Hospital of St Louis St Louis, MO 63110, USA 2Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine 216 South Kingshighway, St Louis, MO 63110, USA

*To whom correspondence should be addressed at: Department of Medicine, The Jewish Hospital of St Louis and Washington University School of Medicine, 216 South Kingshighway, St Louis, MO 63110, USA

Received August 30, 1993. Accepted January 6, 1994.

The platelet-derived growth factor (PDGF) A-chain gene is expressed in a tissue- and developmental stage-specific manner. Here we identify an S1 nuclease sensitive region within the first intron that functions as a negative regulatory element In HeLa but not In human glloblastoma (A172) cells in transient transfection assays. A 147 bp DNA fragment that contains this element functions in a position and orientation independent manner to negatively regulate both the PDGF A-chain promoter and the heterologous herpes simplex virus thymidine kinase (TK) promoter. The cell-type specific effect of this 147 bp DNA fragment is seen when it Is located downstream but not upstream of the reporter gene driven by either the PDGF A-chaln or TK promoters. The negative regulatory element has been localized to a 24 bp DNA sequence within the S1 sensitive site that retains negative regulatory activity and recognizes a nuclear protein in HeLa but not in A172 cells. Furthermore, the 24 bp element functions as a cell type-specific negative element independent of its position. These results suggest that a functional silencer within the first intron exhibits a non-B-form DNA structure under superhelical stress in vitro and may contribute to the cell type-specific transcriptional regulation of PDGF A-chain gene in vivo.

+Present address: Department of Internal Medicine, Research Institute for Nuclear Medicine and Biology, Hiroshima University, 1-2-3 Kasumi, Minami-ku Hiroshima 734, Japan


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