Identification of a negative regulatory site in the upstream region of bovine growth hormone gene

The 5' upstream region of bovine growth hormone (bGH) gene was analyzed. When the region between nucleotides -336 and -240 was deleted, the expression of the reporter chloramphenicol acetyltransferase (CAT) gene increased about 5 fold in HeLa cells and 2.2 fold in rat GH, cells which produce growth hormone. This region was named negative regulatory site (NRS). When NRS was inserted in front of promoter for rat growth hormone (rGH) gene or human thymidine kinase (TK) gene, the CAT activity decreased by 75-80% in HeLa cells and 5-30% in GH, cells. It also repressed the expression of CAT gene from several promoter-enhancer combinations tested. By fine deletion analysis negative elements in the NRS were mapped and found to contain sequences similar to the binding elements of YY1. (C) 1996 Academic Press, Inc.