cis ‐Regulatory analysis for later phase of anterior neuroectoderm‐specific foxQ2 expression in sea urchin embryos
cis ‐Regulatory analysis for later phase of anterior neuroectoderm‐specific foxQ2 expression in sea urchin embryos
复制标题
海胆胚胎前神经外胚层特异性foxQ2表达后期的顺式调控分析
DOI：
10.1002/dvg.23302
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发表时间：
2019
期刊：
影响因子：
1.5
通讯作者：
Yaguchi Shunsuke
中科院分区：
文献类型：
--
作者：
Yamazaki Atsuko;Yamamoto Akane;Yaguchi Junko;Yaguchi Shunsuke
在双边动物中，前神经外胚层的发育是由一个高度保守的分子机制控制的。叉头家族基因foxQ2是已知的关键调控因子之一，在胚胎发生过程中，该基因在该区域合子表达。然而，究竟是什么控制着这一基本因子的表达至今仍不清楚。为了揭示foxq2的调控机制，我们在海胆hemicentrus pulcherrimus中对foxQ2aandfoxQ2b两个foxq2基因进行了调控分析。在海胆胚胎中，foxq2最初在整个动物半球表达，随后在前极区域狭窄表达。在本研究中，作为了解foxq2调控的第一步，我们将重点放在后期的限制性表达上，并利用短半衰期绿色荧光蛋白融合的构建体分析了foxq2a&foxq2bby的上游顺式调控序列。通过对这两个foxq2基因的缺失和突变分析，我们确定了5个长度为4-9 bp的调控序列。这两个调控序列都不包含任何异位激活或空间抑制的基序，这表明后来的mRNA定位是原位调控的。我们还认为，三个氨基酸环延伸类同源盒基因Meis参与了oxq2b的维持，其表达在胚胎发生过程中占主导地位。
The specification of anterior neuroectoderm is controlled by a highly conserved molecular mechanism in bilaterians. A forkhead family gene,foxQ2, is known to be one of the pivotal regulators, which is zygotically expressed in this region during embryogenesis of a broad range of bilaterians. However, what controls the expression of this essential factor has remained unclear to date. To reveal the regulatory mechanism offoxQ2, we performedcis‐regulatory analysis of twofoxQ2genes,foxQ2aandfoxQ2b, in a sea urchinHemicentrotus pulcherrimus. In sea urchin embryos,foxQ2is initially expressed in the entire animal hemisphere and subsequently shows narrower expression restricted to the anterior pole region. In this study, as a first step to understand thefoxQ2regulation, we focused on the later restricted expression and analyzed the upstreamcis‐regulatory sequences offoxQ2aandfoxQ2bby using the constructs fused to short half‐life green fluorescent protein. Based on deletion and mutation analyses of bothfoxQ2, we identified each of the five regulatory sequences, which were 4–9 bp long. Neither of the regulatory sequences contains any motifs for ectopic activation or spatial repression, suggesting that later mRNA localization is regulatedin situ. We also suggest that the three amino acid loop extension‐class homeobox gene Meis is involved in the maintenance offoxQ2b, the expression of which is dominant during embryogenesis.