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DamID 转录谱分析鉴定出 Snail/Scratch 转录因子 Kahuli 是果蝇内脏中胚层中 Alk 的靶标。

DamID transcriptional profiling identifies the Snail/Scratch transcription factor Kahuli as an Alk target in the Drosophila visceral mesoderm.

机构信息

Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, University of Gothenburg, SE-405 30 Gothenburg, Sweden.

Department of Molecular Biology, Umeå University, SE-901 87 Umeå, Sweden.

出版信息

Development. 2021 Dec 1;148(23). doi: 10.1242/dev.199465. Epub 2021 Dec 14.

Abstract

Development of the Drosophila visceral muscle depends on Anaplastic Lymphoma Kinase (Alk) receptor tyrosine kinase (RTK) signaling, which specifies founder cells (FCs) in the circular visceral mesoderm (VM). Although Alk activation by its ligand Jelly Belly (Jeb) is well characterized, few target molecules have been identified. Here, we used targeted DamID (TaDa) to identify Alk targets in embryos overexpressing Jeb versus embryos with abrogated Alk activity, revealing differentially expressed genes, including the Snail/Scratch family transcription factor Kahuli (Kah). We confirmed Kah mRNA and protein expression in the VM, and identified midgut constriction defects in Kah mutants similar to those of pointed (pnt). ChIP and RNA-Seq data analysis defined a Kah target-binding site similar to that of Snail, and identified a set of common target genes putatively regulated by Kah and Pnt during midgut constriction. Taken together, we report a rich dataset of Alk-responsive loci in the embryonic VM and functionally characterize the role of Kah in the regulation of embryonic midgut morphogenesis.

摘要

果蝇内脏肌肉的发育依赖于 Anaplastic Lymphoma Kinase(Alk)受体酪氨酸激酶(RTK)信号通路,该通路特异性地指定了圆形内脏中胚层(VM)中的创始细胞(FCs)。尽管其配体 Jelly Belly(Jeb)激活 Alk 已得到很好的描述,但很少有靶分子被鉴定出来。在这里,我们使用靶向 DamID(TaDa)技术,在过表达 Jeb 的胚胎和 Alk 活性被阻断的胚胎中鉴定 Alk 的靶标,揭示了差异表达的基因,包括 Snail/Scratch 家族转录因子 Kahuli(Kah)。我们在 VM 中证实了 Kah mRNA 和蛋白的表达,并在 Kah 突变体中鉴定出类似于 pointed(pnt)的中肠收缩缺陷。ChIP 和 RNA-Seq 数据分析定义了一个与 Snail 相似的 Kah 靶标结合位点,并鉴定了一组可能受 Kah 和 Pnt 调节的共同靶基因,它们在中肠收缩过程中发挥作用。综上所述,我们报告了胚胎 VM 中 Alk 反应性基因座的丰富数据集,并对 Kah 在调节胚胎中肠形态发生中的作用进行了功能表征。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8565/8722224/fe680d4103d6/develop-148-199465-g1.jpg

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