Complex interactions between cis-regulatory modules in native conformation are critical for Drosophila snail expression
Complex interactions between cis-regulatory modules in native conformation are critical for Drosophila snail expression
复制标题
DOI：
10.1242/dev.069146
复制
发表时间：
2011-09-15
期刊：
影响因子：
4.6
通讯作者：
Stathopoulos, Angelike
中科院分区：
文献类型：
--
作者：
Dunipace, Leslie;Ozdemir, Anil;Stathopoulos, Angelike
它已被证明在几种生物体中，多个顺式调控模块（CRM）的基因座可以同时激活，以支持类似的时空表达。为了了解这种看似多余的标准物质的功能的重要性，我们研究了两个标准物质从果蝇蜗牛基因位点，这两个都是活跃的腹侧区域的前原肠胚。通过使用BAC重组工程和定点转基因在类似于25 kb DNA拯救构建体中进行一系列缺失，我们证明了这两个CRM不是冗余的。远端CRM是绝对需要的生存能力，而近端CRM仅在极端条件下需要，如高温。与其不同的要求一致，CRM支持不同的表达模式：近端CRM表现出相对于内源性蜗牛扩展的表达域，而远端CRM表现出几乎完全重叠，除了在最前极与蜗牛。我们进一步表明，远端CRM通常限制了近端CRM的表达域增加，并且近端CRM充当由远端CRM驱动的表达水平的“阻尼器”。因此，这两个CRM相互作用的顺式在一个非加性的方式，这些相互作用可能是重要的微调的结构域和基因表达水平。
It has been shown in several organisms that multiple cis-regulatory modules (CRMs) of a gene locus can be active concurrently to support similar spatiotemporal expression. To understand the functional importance of such seemingly redundant CRMs, we examined two CRMs from the Drosophila snail gene locus, which are both active in the ventral region of pre-gastrulation embryos. By performing a deletion series in a similar to 25 kb DNA rescue construct using BAC recombineering and site-directed transgenesis, we demonstrate that the two CRMs are not redundant. The distal CRM is absolutely required for viability, whereas the proximal CRM is required only under extreme conditions such as high temperature. Consistent with their distinct requirements, the CRMs support distinct expression patterns: the proximal CRM exhibits an expanded expression domain relative to endogenous snail, whereas the distal CRM exhibits almost complete overlap with snail except at the anterior-most pole. We further show that the distal CRM normally limits the increased expression domain of the proximal CRM and that the proximal CRM serves as a 'damper' for the expression levels driven by the distal CRM. Thus, the two CRMs interact in cis in a non-additive fashion and these interactions may be important for fine-tuning the domains and levels of gene expression.