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睾丸特异性组蛋白H1t基因的表达:多个顺式作用启动子元件参与的证据。

Expression of the testis-specific histone H1t gene: evidence for involvement of multiple cis-acting promoter elements.

作者信息

Wolfe S A, van Wert J M, Grimes S R

机构信息

Research Service (151), Overton Brooks Veterans Administration Medical Center, Shreveport, Louisiana 71101-4295, USA.

出版信息

Biochemistry. 1995 Sep 26;34(38):12461-9. doi: 10.1021/bi00038a045.

Abstract

The histone H1t gene is expressed exclusively in testis primary spermatocytes. Previous studies indicate that accumulation of H1t mRNA occurs only in primary spermatocytes in normal rats and in transgenic mice bearing the rat H1t transgene. In this study, DNA sequences of human, monkey, mouse, and rat H1t genes were compared and found to be almost identical in the proximal promoter region extending from the H1/AC box through the TATAA box. In addition to conserved elements common to replication-dependent H1 promoters, the H1t promoter contains a unique TE element, and sequences within this element may contribute to enhanced expression of the gene in primary spermatocytes. Two imperfect inverted repeat sequences designated TE1 and TE2, that are located within the larger TE element, overlap a central GC-rich region and bind specifically to nuclear proteins derived from primary spermatocytes. Protein interactions characterized by methylation interference and UV cross-linking experiments indicate that a complex of proteins with a molecular mass of approximately 180 kDa binds TE1. The GC-rich region in H1t and in some replication dependent histone H1 promoters contains an Sp1 consensus sequence. Although the H1t/TE element that contains the GC-rich region binds nuclear proteins, it does not appear to bind Sp1 obtained from cell populations enriched in primary spermatocytes as determined by electrophoretic mobility supershift assays using polyclonal anti-Sp1 antibodies.

摘要

组蛋白H1t基因仅在睾丸初级精母细胞中表达。先前的研究表明,H1t mRNA的积累仅发生在正常大鼠和携带大鼠H1t转基因的转基因小鼠的初级精母细胞中。在本研究中,对人、猴、小鼠和大鼠H1t基因的DNA序列进行了比较,发现从H1/AC盒到TATAA盒的近端启动子区域几乎相同。除了依赖复制的H1启动子共有的保守元件外,H1t启动子还包含一个独特的TE元件,该元件内的序列可能有助于该基因在初级精母细胞中的增强表达。位于较大TE元件内的两个不完美反向重复序列TE1和TE2,与富含GC的中央区域重叠,并特异性结合来自初级精母细胞的核蛋白。通过甲基化干扰和紫外线交联实验表征的蛋白质相互作用表明,一种分子量约为180 kDa的蛋白质复合物结合TE1。H1t和一些依赖复制的组蛋白H1启动子中的富含GC的区域包含一个Sp1共有序列。尽管包含富含GC区域的H1t/TE元件结合核蛋白,但通过使用多克隆抗Sp1抗体的电泳迁移率超迁移分析确定,它似乎不结合从富含初级精母细胞的细胞群体中获得的Sp1。

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