Histone gene expression and histone mRNA 3' end structure in Caenorhabditis elegans.
Histone gene expression and histone mRNA 3' end structure in Caenorhabditis elegans.
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DOI：
10.1186/1471-2199-8-51
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发表时间：
2007-06-14
影响因子：
--
通讯作者：
Müller B
中科院分区：
文献类型：
--
作者：
Keall R;Whitelaw S;Pettitt J;Müller B
组蛋白的合成是细胞增殖和DNA包装成染色质所必需的。在动物中，组蛋白是由多拷贝复制依赖性组蛋白基因的表达提供的。组蛋白mrna通过组蛋白特异性机制加工，在高度保守的RNA发夹元件后结束，并且缺乏poly(a)尾巴。在脊椎动物和果蝇中，它们的表达依赖于与RNA发夹元件结合的HBP/SLBP。我们之前表明，这些顺式和反式的组蛋白基因表达调节因子在秀丽隐杆线虫中是保守的。本文报道了线虫发育过程中组蛋白mRNA 3'端结构和组蛋白基因表达的研究结果。复制依赖性组蛋白基因的序列分析显示，在组蛋白前mrna的3'非翻译区存在几个高度保守的序列元件，包括RNA发夹元件和多聚腺苷化信号。为了确定秀丽隐杆线虫组蛋白mRNA的3′端形成是否发生在这种多腺苷化信号中并导致组蛋白mRNA的多腺苷化，我们研究了组蛋白mRNA的mRNA 3′端结构。通过对组蛋白mRNA末端的poly(A)选择、RNAse保护和测序，我们确定了大多数线虫组蛋白mRNA缺乏poly(A)尾巴，在发夹结构后3到6个核苷酸处结束，在A或U之后，并且具有3' OH基团。RNAi敲低秀丽隐杆线虫HBP/SLBP的CDL-1对组蛋白mRNA水平没有显著影响，但会严重消耗组蛋白水平。组蛋白基因的表达在发育过程中有所不同，与L1动物和成年动物相比，L3动物的组蛋白基因表达减少。在成人中，组蛋白基因的表达仅限于细胞分裂发生的生殖系。我们的研究结果表明，秀丽隐杆线虫组蛋白基因的表达受到与其他动物相似的控制机制：秀丽隐杆线虫组蛋白mRNA 3‘端的结构与组蛋白特异性mRNA 3’端的加工是兼容的；CDL-1在组蛋白基因转录后调控中的作用秀丽隐杆线虫组蛋白mRNA水平在细胞分裂活跃时期升高，表明组蛋白基因表达与DNA复制有关。
Histone protein synthesis is essential for cell proliferation and required for the packaging of DNA into chromatin. In animals, histone proteins are provided by the expression of multicopy replication-dependent histone genes. Histone mRNAs that are processed by a histone-specific mechanism to end after a highly conserved RNA hairpin element, and lack a poly(A) tail. In vertebrates and Drosophila, their expression is dependent on HBP/SLBP that binds to the RNA hairpin element. We showed previously that these cis and trans acting regulators of histone gene expression are conserved in C. elegans. Here we report the results of an investigation of the histone mRNA 3' end structure and of histone gene expression during C. elegans development. Sequence analysis of replication-dependent histone genes revealed the presence of several highly conserved sequence elements in the 3' untranslated region of histone pre-mRNAs, including an RNA hairpin element and a polyadenylation signal. To determine whether in C. elegans histone mRNA 3' end formation occurs at this polyadenylation signal and results in polyadenylated histone mRNA, we investigated the mRNA 3' end structure of histone mRNA. Using poly(A) selection, RNAse protection and sequencing of histone mRNA ends, we determined that a majority of C. elegans histone mRNAs lack a poly(A) tail and end three to six nucleotides after the hairpin structure, after an A or a U, and have a 3' OH group. RNAi knock down of CDL-1, the C. elegans HBP/SLBP, does not significantly affect histone mRNA levels but severely depletes histone protein levels. Histone gene expression varies during development and is reduced in L3 animals compared to L1 animals and adults. In adults, histone gene expression is restricted to the germ line, where cell division occurs. Our findings indicate that the expression of C. elegans histone genes is subject to control mechanisms similar to the ones in other animals: the structure of C. elegans histone mRNA 3' ends is compatible with histone-specific mRNA 3' end processing; CDL-1 functions in post-transcriptional control of histone gene expression; and C. elegans histone mRNA levels are elevated at periods of active cell division, indicating that histone gene expression is linked to DNA replication.
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