Co-regulation of invected and engrailed by a complex array of regulatory sequences in Drosophila.
Co-regulation of invected and engrailed by a complex array of regulatory sequences in Drosophila.
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DOI：
10.1016/j.ydbio.2014.08.021
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发表时间：
2014-11-01
影响因子：
2.7
通讯作者：
Kassis, Judith A.
中科院分区：
文献类型：
--
作者：
Cheng, Yuzhong;Brunner, Alayne L.;Kremer, Stefanie;DeVido, Sarah K.;Stefaniuk, Catherine M.;Kassis, Judith A.
Invected （inv）和engrailed （en）形成了一个长约115kb的基因复合体。这两个基因编码高度相关的同源结构域蛋白，在整个发育过程中以复杂的方式共同调节。我们对inv/en调控DNA的解剖表明，大多数增强子分布在一个62kb的区域。我们使用了两种类型的构建体来分析该DNA的功能：基于p元素的报告基因构建体，将小片段的DNA融合到驱动lacZ表达的en启动子上，以及使用phiC31系统将ha标记的en和inv插入基因组中的大构建体。此外，我们在原位产生了inv和en DNA的缺失，并测定了它们对inv/en表达的影响。我们的研究结果支持并扩展了我们在创新/环保法规方面的知识。首先，inv和en共享调控DNA，其中大部分位于en转录单元的两侧。为了支持这一点，一个79 kb的HA-en转基因可以拯救未被发现的双突变体到有活力的、可生育的成年体。相反，一个84kb的HA-inv转基因缺乏inv/en表达的大部分增强子。第二，胚胎中有多种inv/en条纹增强子；其中一些可能是多余的，但其他的在胚胎发育的不同阶段起着独立的作用。最后，没有小的报告结构在影像椎间盘后腔室表达，这是无/无表达的标志。在79 kb的HA-en转基因中，HA-en在影像椎间盘后腔室的表达明显增强，而在45 kb的HA-en转基因中，HA-en在影像椎间盘的表达较弱且可变。我们认为，成像光盘增强子的活性取决于inv/en结构域的染色质结构。
invected (inv) and engrailed (en) form a gene complex that extends about 115kb. These two genes encode highly related homeodomain proteins that are co-regulated in a complex manner throughout development. Our dissection of inv/en regulatory DNA shows that most enhancers are spread throughout a 62kb region. We used two types of constructs to analyze the function of this DNA: P-element based reporter constructs with small pieces of DNA fused to the en promoter driving lacZ expression and large constructs with HA-tagged en and inv inserted in the genome with the phiC31 system. In addition, we generated deletions of inv and en DNA in situ and assayed their effects on inv/en expression. Our results support and extend our knowledge of inv/en regulation. First, inv and en share regulatory DNA, most of which is flanking the en transcription unit. In support of this, a 79-kb HA-en transgene can rescue inv en double mutants to viable, fertile adults. In contrast, an 84-kb HA-inv transgene lacks most of the enhancers for inv/en expression. Second, there are multiple enhancers for inv/en stripes in embryos; some of these may be redundant but others play discrete roles at different stages of embryonic development. Finally, no small reporter construct gave expression in the posterior compartment of imaginal discs, a hallmark of inv/en expression. Robust expression of HA-en in the posterior compartment of imaginal discs is evident from the 79-kb HA-en transgene, while a 45-kb HA-en transgene gives weaker, variable imaginal disc expression. We suggest that the activity of the imaginal disc enhancer(s) is dependent on the chromatin structure of the inv/en domain.
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影响因子：
4.6
作者：
Kwon, Deborah;Mucci, Diane;Kassis, Judith A.
通讯作者：
Kassis, Judith A.
影响因子：
64.8
作者：
通讯作者：
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影响因子：
64.8
作者：
DINARDO, S;SHER, E;OFARRELL, PH
通讯作者：
OFARRELL, PH
影响因子：
4
作者：
Barolo, Scott
通讯作者：
Barolo, Scott
影响因子：
64.5
作者：
Junion, Guillaume;Spivakov, Mikhail;Furlong, Eileen E. M.
通讯作者：
Furlong, Eileen E. M.