| Popis: |
Analysis of the chromatin of different cell types has identified four DNase I-hypersensitive sites in the 5′-flanking region of the α1(VI) collagen gene, mapping at −4.6, −4.4, −2.5, and −0.1 kilobase (kb) from the RNA start site. The site at −2.5 kb was independent from, whereas the other three sites could be related to, α1(VI) mRNA expression. The site at −0.1 kb was present in cells expressing (NIH3T3 and C2C12) but absent in cells not expressing (EL4) the mRNA; the remaining two sites were apparently related with high levels of mRNA. DNase I footprinting and gel-shift assays with NIH3T3 and C2C12 nuclear extracts have located a binding site for transcription factor AP1 (activator protein 1) between nucleotides −104 and −73. When nuclear extracts from EL4 lymphocytes were used, the AP1 site-containing sequence was bound by proteins not related to AP1. The existence of the hypersensitive site at −0.1 kb may be related to the binding of AP1 and of additional factors to the core promoter (Piccolo, S., Bonaldo, P., Vitale, P., Volpin, D., and Bressan, G. M. (1995) J. Biol. Chem. 270, 19583–19590). The function of the AP1 binding site and of the core promoter in the transcriptional regulation of the Col6a1gene was investigated by expressing several promoter-reporter gene constructs in transgenic mice and in cell cultures. The results indicate that regulation of transcription of the Col6a1gene by different cis-acting elements (core promoter, AP1 binding site and enhancers) is not completely modular, but the final output depends on the specific interactions among the three elements in a defined cell type. |
