DEVELOPMENTAL CONTROL OF PROMOTER-SPECIFIC FACTORS RESPONSIBLE FOR THE EMBRYONIC ACTIVATION AND INACTIVATION OF THE SEA-URCHIN EARLY HISTONE H-3 GENE
DEVELOPMENTAL CONTROL OF PROMOTER-SPECIFIC FACTORS RESPONSIBLE FOR THE EMBRYONIC ACTIVATION AND INACTIVATION OF THE SEA-URCHIN EARLY HISTONE H-3 GENE
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DOI：
10.1101/gad.3.7.973
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发表时间：
1989-07-01
影响因子：
10.5
通讯作者：
CHILDS, G
中科院分区：
文献类型：
--
作者：
DILIBERTO, M;LAI, ZC;CHILDS, G
我们已经开始了对紫癜海胆早期组蛋白H3基因的时间胚胎表达的分子基础的研究。克隆的构建体在显微注射到相关海胆物种Lytechinus pictus的单细胞受精卵后显示出适当的时间调节。H3基因上游启动子区域的缺失分析揭示了几个参与阳性和阴性对照的区域，dna酶I足迹，迁移转移和甲基化干扰实验揭示了多个序列特异性dna结合蛋白，这些蛋白与RNA起始位点上游200 bp内至少五个不同的区域相互作用。从分期胚胎中提取的提取物显示，这些因子结合其靶序列的能力受到调节。只有在H3基因转录活跃的阶段，才能检测到在四个不同位点结合的蛋白质。此外，三种不同形式的ccaat结合蛋白也受到暂时调节。然而，这些蛋白的活性与基因的转录活性呈负相关。TATA box和CCAAT序列是表达低水平H3转录本所需的全部序列，具有适当的时间模式。这种方法应该有助于理解在早期胚胎发生过程中用于调节基因表达时间模式的机制。
We have begun an investigation of the molecular basis for the temporal embryonic expression of the early histone H3 gene of the sea urchin Stronglyocentrotus purpuratus. Cloned constructs exhibit the proper temporal regulation following microinjection into one-cell zygotes of the related sea urchin species, Lytechinus pictus. Deletion analysis of the upstream promoter region of the H3 gene revealed several regions that are involved in both positive and negative control DNase I footprinting, mobility shift, and methylation interference experiments reveal multiple sequence-specific DNA-binding proteins that interact with at least five distinct regions within 200 bp upstream of the RNA initiation site. Extracts prepared from staged embryos revealed that the ability of the factors to bind their target sequences was regulated. Proteins bound at four different sites were detected only at stages when the H3 gene was active transcriptionally. In addition, three different forms of a CCAAT-binding protein also are regulated temporally. The activity of these protein(s), however, correlates inversely within the transcriptional activity of the gene. The TATA box and CCAAT sequences are all that is required for expression of low levels of H3 transcripts with the proper temporal pattern. This approach should be useful in understanding the mechanisms used to regulate temporal patterns of gene expression during early embryogenesis.