Structural and functional analysis of the rat testis-specific histone H1t gene.
Structural and functional analysis of the rat testis-specific histone H1t gene.
复制标题
大鼠睾丸特异性组蛋白 H1t 基因的结构和功能分析。
DOI：
10.1002/jcb.240440102
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发表时间：
1990
影响因子：
4
通讯作者：
Stein,JL
中科院分区：
文献类型：
--
作者：
Grimes,SR;Wolfe,SA;Anderson,JV;Stein,GS;Stein,JL
已对含有睾丸特异性组蛋白H1 t基因和组蛋白H4 t基因的6.86 kb大鼠基因组DNA片段进行了测序。对大鼠肝脏和睾丸总细胞RNA的S1核酸酶保护分析表明，组蛋白H1 t mRNA仅存在于睾丸中。各种高度富集的大鼠睾丸细胞类型的人群的检查显示，H1 t mRNA被发现完全在粗线期精母细胞中富集的馏分。当蛋白质，DNA相互作用的近端启动子区域内的组蛋白H1 t基因进行了研究，电泳迁移率变动分析，只有微小的差异被发现在迁移率变动模式的H1 t启动子的测定比较结合的核蛋白从粗线期精母细胞和早期精子细胞。然而，在比较大鼠粗线期精母细胞与肝脏的核蛋白时，观察到结合的主要差异。比较大鼠睾丸、大鼠肝癌H4细胞、HeLa细胞和COS-1细胞的结合模式也显示出显著差异。通过在转染的大鼠肝癌H4细胞、HeLa细胞和COS-1细胞中的瞬时表达测定中测量H1 t促进的氯霉素乙酰转移酶（CAT）mRNA水平来检查组蛋白H1 t启动子的转录活性。这些试验表明，与用亲本SV 40促进的载体pSV 2CAT表达相比，组蛋白H1 t促进的CAT基因在HeLa细胞和COS-1细胞中的功能较差。H1 t启动的CAT基因在转染的大鼠肝癌H4细胞中明显不起作用，这与组蛋白H1 t基因在睾丸生殖细胞中的特异性表达相一致。
A 6.86 kb rat genomic DNA fragment containing the testis‐specific histone H1t gene and the histone H4t gene has been sequenced. S1‐nuclease protection analyses of total cellular RNA from rat liver and testis showed that histone H1t mRNA was present only in testis. Examination of various highly enriched populations of rat testis cell types revealed that H1t mRNA was found exclusively in a fraction enriched in pachytene spermatocytes. When protein, DNA interactions within the proximal promoter region of the histone H1t gene were examined by electrophoretic mobility shift assays, only minor differences were found in mobility shift patterns of the H1t promoter in assays comparing binding of nuclear proteins from pachytene spermatocytes and early spermatids. However, major differences in binding were observed upon comparing nuclear proteins from rat pachytene spermatocytes to liver. Comparison of binding patterns of rat testis, rat hepatoma H4 cells, HeLa cells, and COS‐1 cells also revealed dramatic differences. Transcriptional activity of the histone H1t promoter was examined by measuring H1t promoted chloramphenicol acetyltransferase (CAT) mRNA levels in transient experession assays in transfected rat hepatoma H4 cells, HeLa cells, and COS‐1 cells. These assays revealed that the histone H1t promoted CAT gene functioned poorly in HeLa cells and COS‐1 cells compared to expression with the parent SV40 promoted vector pSV2CAT. The H1t promoted CAT gene apparently did not work at all in transfected rat hepatoma H4 cells, which is consistent with testis germinal cell specific expression of the histone H1t gene.