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首页> 外文期刊>Molecular and Cellular Biology >Sea urchin early and late H4 histone genes bind a specific transcription factor in a stable preinitiation complex.
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Sea urchin early and late H4 histone genes bind a specific transcription factor in a stable preinitiation complex.

机译：海胆早期和晚期H4组蛋白基因在稳定的预启动复合物中结合特定的转录因子。

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摘要

Early embryonic H4 (EH4) and H2B (EH2B) and late embryonic H4 (LH4) histone genes were transcribed in vitro in a nuclear extract from hatching blastula embryos of the sea urchin Strongylocentrotus purpuratus. The extract was prepared by slight modifications of the methods of Morris et al. (G. F. Morris, D. H. Price, and W. F. Marzluff, Proc. Natl. Acad. Sci. USA 83:3674-3678, 1986) that have been used to obtain a cell-free transcription system from embryos of the sea urchin Lytechinus variegatus. Achievement of maximum levels of transcription of the EH4 and LH4 genes required a 5- to 10-min preincubation of template with extract in the absence of ribonucleoside triphosphates. This preincubation allowed the formation of a stable complex which was preferentially transcribed compared with a second EH4 or LH4 template that was added 10 min later. Although the EH4 gene inhibited both EH4 and LH4 gene transcription in this assay and although the LH4 gene inhibited both EH4 and LH4 genes, neither of these genes inhibited transcription of the EH2B gene. Preincubation with the EH2B gene had no effect on the transcription of subsequently added EH4 or LH4 genes. Using this template commitment assay, we showed that the site of binding of at least one essential factor required for transcription of both EH4 and LH4 genes was located between positions -102 and -436 relative to the 5' terminus of the EH4 mRNA. Moreover, deletion of this region resulted in a reduction in EH4 gene transcription in vitro. The sea urchin gene-specific trans-acting factors, in the analysis of the cis-acting sequences with which they interact, and in biochemical studies on the formation of stable transcription complexes.
机译：早期胚胎H4（EH4）和H2B（EH2B）和晚期胚胎H4（LH4）组蛋白基因在海胆Strongylocentrotus purpuratusus孵化的囊胚胚胎的核提取物中体外转录。提取物是通过稍稍修改Morris等人的方法制备的。 （G.F.Morris，D.H.Price，和W.F.Marzluff，Proc.Natl.Acad.Sci.USA 83：3674-3678，1986）已经用于从海胆Lytechinus variegatus的胚胎中获得无细胞的转录系统。要实现EH4和LH4基因最大转录水平，需要在没有三磷酸核糖核苷的情况下，用提取物对模板进行5至10分钟的预孵育。与在10分钟后添加的第二个EH4或LH4模板相比，该预孵育允许形成稳定的复合物，其优先被转录。尽管在该测定中，EH4基因同时抑制了EH4和LH4基因的转录，尽管LH4基因同时抑制了EH4和LH4基因，但这些基因均未抑制EH2B基因的转录。与EH2B基因预孵育对随后添加的EH4或LH4基因的转录没有影响。使用该模板承诺分析，我们显示了EH4和LH4基因转录所需的至少一种必需因子的结合位点位于相对于EH4 mRNA 5'端的位置-102和-436之间。而且，该区域的缺失导致体外EH4基因转录的减少。海胆基因特异的反式作用因子，在与它们相互作用的顺式作用序列的分析中，以及在有关稳定转录复合物形成的生化研究中。

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