Ashe H L, Monks J, Wijgerde M, Fraser P, Proudfoot N J
Sir William Dunn School of Pathology, University of Oxford, UK.
Genes Dev. 1997 Oct 1;11(19):2494-509. doi: 10.1101/gad.11.19.2494.
We have identified novel nuclear transcripts in the human beta-globin locus using nuclear run-on analysis in erythroid cell lines and in situ hybridization analysis of erythroid tissue. These transcripts extend across the LCR and intergenic regions but are undetectable in nonerythroid cells. Surprisingly, transient transfection of a beta-globin gene (epsilon, gamma, or beta) induces transcription of the LCR and intergenic regions from the chromosomal beta-globin locus in nonerythroid cell lines. The beta-globin genes themselves, however, remain transcriptionally silent. Induction is dependent on transcription of the globin gene in the transfected plasmid but does not require protein expression. Using in situ hybridization analysis, we show that the plasmid colocalizes with the endogenous beta-globin locus providing insight into the mechanism of transinduction.
我们通过在红系细胞系中进行核转录分析以及对红系组织进行原位杂交分析,在人类β-珠蛋白基因座中鉴定出了新的核转录本。这些转录本跨越了LCR和基因间区域,但在非红系细胞中无法检测到。令人惊讶的是,β-珠蛋白基因(ε、γ或β)的瞬时转染可诱导非红系细胞系中染色体β-珠蛋白基因座的LCR和基因间区域的转录。然而,β-珠蛋白基因本身仍保持转录沉默。诱导作用依赖于转染质粒中珠蛋白基因的转录,但不需要蛋白质表达。通过原位杂交分析,我们表明质粒与内源性β-珠蛋白基因座共定位,这为反式诱导机制提供了深入了解。