Role of chromatin structure in transcriptional regulation of MMTV LTR hormone-dependent promoter.
Abstract
Studies of chromatin structure were performed in mouse fibroblast cell lines containing Bovine Papilloma Virus (BPV) based artificial minichromosomes containing Mouse Mammary Tumor Virus (MMTV) Long Terminal Repeat (LTR), a retroviral promoter regulated by glucocorticoids, driving the transcription of v-Ha-ras. These minichromosomes fractionate with the "active chromatin", indicating an association of the minichromosomes with components of the "nuclear matrix". Two regions of the minichromosomes upstream and downstream of v-Ha-ras are involved in this interaction. MMTV LTR promoter is associated with nucleosomes precisely positioned on the DNA sequences. Hormonal activation is accompanied by a structural change of the nucleosome associated with the hormone response elements (HREs). This structural change can be visualized by the appearance of a hormono-dependent DNaseI hypersensitive site. Anti-hormones, even when able to promote a strong binding of the receptor to the nucleus, are unable to induce the chromatin structural change. The strong association of the hormone-receptor complex with the nucleus is necessary to induce the DNaseI hypersensitive site and to maintain the transcription, but is not necessary for DNaseI hypersensitivity maintenance. This suggests a double role for the hormone-receptor complex: 1) induction of a chromatin rearrangement and 2) transcriptional transactivation.