Specific binding of nuclear proteins to a bifunctional promoter element upstream of the H1/AC box of the testis-specific histone H1t gene.
Specific binding of nuclear proteins to a bifunctional promoter element upstream of the H1/AC box of the testis-specific histone H1t gene.
复制标题
核蛋白与睾丸特异性组蛋白 H1t 基因 H1/AC 盒上游的双功能启动子元件特异性结合。
DOI：
10.1095/biolreprod.102.014084
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Grimes,SidneyR
中科院分区：
文献类型：
--
作者：
Wolfe,StevenA;Grimes,SidneyR
睾丸特异性组蛋白H1 t基因在精子发生过程中只在初级精母细胞中转录。对转基因小鼠的研究表明，H1 t近端启动子的141个碱基对（bp）与800 bp的下游序列相伴随足以进行组织特异性转录。来自睾丸和粗线期精母细胞的核蛋白产生跨越覆盖H1 t转录起始位点上游100至125个核苷酸的阻遏元件（RE）的区域的足迹。在电泳迁移率变动分析中，只有睾丸核蛋白与该元件的5′端结合并产生独特的低迁移率复合物。这种睾丸复合物与由来自几种细胞系的阻遏蛋白形成的复合物不同，阻遏蛋白与元件的3′端结合。当使用来自初级精母细胞的核蛋白时，形成睾丸复合体条带，其中H1 t基因被转录，并且当使用来自早期精子细胞的核蛋白时，条带强度下降70%-80%，其中H1 t基因转录停止。使用睾丸核蛋白的蛋白质-DNA交联实验在SDS/PAGE凝胶上产生59、52和50 kDa的电泳条带。
The testis-specific histone H1t gene is transcribed exclusively in primary spermatocytes during spermatogenesis. Studies with transgenic mice show that 141 base pairs (bp) of the H1t proximal promoter accompanied with 800 bp of downstream sequence are sufficient for tissue-specific transcription. Nuclear proteins from testis and pachytene spermatocytes produce footprints spanning the region covering the repressor element (RE) from 100 to 125 nucleotides upstream of the H1t transcriptional initiation site. Only testis nuclear proteins bind to the 5′-end of the element and produce a unique, low-mobility complex in electrophoretic mobility shift assays. This testis complex is distinct from the complex formed by a repressor protein derived from several cell lines that binds to the 3′-end of the element. The testis complex band is formed when using nuclear proteins from primary spermatocytes, where the H1t gene is transcribed, and band intensity drops 70%–80% when using nuclear proteins from early spermatids, where H1t gene transcription ceases. Protein-DNA cross-linking experiments using testis nuclear proteins produce electrophoretic bands of 59, 52, and 50 kDa on SDS/PAGE gels.