The essential Drosophila CLAMP protein differentially regulates non-coding roX RNAs in male and females.
The essential Drosophila CLAMP protein differentially regulates non-coding roX RNAs in male and females.
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DOI：
10.1007/s10577-016-9541-9
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发表时间：
2017-06
期刊：
影响因子：
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通讯作者：
Larschan EN
中科院分区：
文献类型：
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作者：
Urban JA;Doherty CA;Jordan WT 3rd;Bliss JE;Feng J;Soruco MM;Rieder LE;Tsiarli MA;Larschan EN
异配物种需要染色体范围的基因调控来补偿性染色体基因剂量的差异。在黑腹果蝇中，通过招募雄性特异性致死 (MSL) 复合体，单个雄性 X 染色体的转录输出与 XX 雌性的转录输出相等，从而使活性基因的转录水平增加两倍。 MSL 复合体包含多种蛋白质成分和两个由 MSL 复合体转录激活的非编码 roX（X 上的 RNA）RNA。我们之前发现 MSL 复合物靶向 X 染色体依赖于 MSL 蛋白染色质连接接头 (CLAMP) 锌指蛋白。为了更好地了解 CLAMP 功能，我们使用 CRISPR/Cas9 基因组编辑系统在钳基因中产生移码突变，从而消除 CLAMP 蛋白的表达。我们发现钳无效雌性在第三龄幼虫阶段死亡，而几乎所有钳无效雄性在早期发育阶段死亡。此外，我们发现在clamp null雌性中roX基因表达被激活，而在clamp null雄性中roX基因表达降低。因此，CLAMP 以性别特异性方式调节 roX 丰度。我们的结果为重要转录因子对性别特异性基因的调控提供了新的见解。
Heterogametic species require chromosome-wide gene regulation to compensate for differences in sex chromosome gene dosage. In Drosophila melanogaster, transcriptional output from the single male X-chromosome is equalized to that of XX females by recruitment of the Male Specific Lethal (MSL) complex, which increases transcript levels of active genes two-fold. MSL complex contains several protein components and two non-coding roX (RNA on the X) RNAs that are transcriptionally activated by MSL complex. We previously discovered that targeting of MSL complex to the X-chromosome is dependent on the Chromatin-Linked Adapter for MSL Protein (CLAMP) zinc finger protein. To better understand CLAMP function, we used the CRISPR/Cas9 genome editing system to generate a frameshift mutation in the clamp gene that eliminates expression of CLAMP protein. We found that clamp null females die at the third instar larval stage, while almost all clamp null males die at earlier developmental stages. Moreover, we found that in clamp null females roX gene expression is activated whereas in clamp null males roX gene expression is reduced. Therefore, CLAMP regulates roX abundance in a sex-specific manner. Our results provide new insights into sex-specific gene regulation by an essential transcription factor.