Significance of the chromatin structure in expression of the rat prolactin gene.

Abstract

To elucidate the mechanisms underlying cell type-specific expression of the growth hormone (GH) and prolactin (PRL) genes, we used rat pituitary-derived cell lines producing exclusively GH (GC cells) or PRL (235 cells), and examined the following: expression of transcription factors essential for GH and/or PRL gene expression; promoter/enhancer activity of the GH and PRL genes transiently introduced by transfection; and chromatin structures of the GH and PRL genes. Even in PRL-nonproducing GC cells, the PRL promoter/enhancer was more active than the GH promoter, and the transcription factors, Pit-1 and estrogen receptor (ER), essential for PRL gene expression were functional. The PRL promoter/enhancer of GC cells was normal. On DNase I sensitivity analysis of the chromatin structure, two hypersensitive sites were detected in PRL gene chromatin of PRL-producing 235 cells but none in that of GC cells. It thus follows that the major reason for absence of the expression of the endogenous PRL gene in GC cells is neither the lack of transcription factors necessary for PRL gene expression nor an anomaly of the PRL gene itself, but that the chromatin structure of the PRL gene differs in PRL-nonproducing and -producing cells. It was shown in this study that neither Pit-1 nor ER is required for conversion of the structure of PRL gene chromatin to a DNase I-hypersensitive state.