Cloning and expression analysis of the chick DAN gene, an antagonist of the BMP family of growth factors
Cloning and expression analysis of the chick DAN gene, an antagonist of the BMP family of growth factors
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DOI：
10.1002/dvdy.10079
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发表时间：
2002-05-01
影响因子：
2.5
通讯作者：
Barald, KF
中科院分区：
文献类型：
--
作者：
Gerlach-Bank, LM;Ellis, AD;Barald, KF
神经母细胞瘤（DAN）中的差异筛选选择性基因异常是胱氨酸结蛋白家族的成员，该家族包括Cerberus和Gremlin。首先在筛选中分离出在转化的大鼠成纤维细胞中下调的基因，随后在非洲爪蟾、小鼠和人类中克隆了DAN。DAN的过表达抑制转化的表型并延迟细胞进入S期。生物化学分析表明，DAN能够结合骨形态发生蛋白并拮抗其信号传导活性。本研究对鸡DAN基因进行了克隆和测序，发现其具有保守的胱氨酸结区和N-糖基化位点。根据鸡DAN基因3'端编码序列设计核糖核酸探针，应用原位杂交技术对鸡胚发育过程中的DAN进行分析。鸡DAN在第10阶段开始在发育中的体节和中耳上皮中表达。在第14阶段，眼睛的神经层中的表达变得明显。到第17阶段，表达已经扩展到后脑的基部。肢芽标记开始于第20阶段，而在鳃弓中的表达出现在第25阶段。比较原位杂交结果显示，鸡DAN表达与小鼠DAN表达基本一致。然而，鸡DAN被发现在耳上皮和脊索，而小鼠DAN仅限于上覆的耳外间充质和缺席的脊索。这一观察结果表明，DAN可能在鸡和小鼠耳和脊索发育中发挥不同的作用。(C)2002 Wiley-Liss，Inc.
Differential screening-selected gene aberrative in neuroblastoma (DAN) is a member of a cystine knot protein family that includes Cerberus and Gremlin. First isolated in a screen to identify genes down-regulated in transformed rat fibroblasts, DAN has subsequently been cloned in Xenopus, mouse, and human. Overexpression of DAN suppresses the transformed phenotype and retards the cell's entry into S phase. Biochemical analyses have demonstrated DAN's ability to bind bone morphogenetic proteins and antagonize their signaling activity. In this study, chick DAN was cloned and sequenced, revealing a conserved cystine knot region as well as an N-glycosylation site. A riboprobe was designed from the 3' chick DAN coding sequence and used for analysis of DAN in the developing chick embryo by in situ hybridization. Chick DAN was expressed beginning at stage 10 in the developing somites and the medial otic epithelium. Expression in the neural layer of the eye became apparent at stage 14. By stage 17, expression had expanded to the base of the hindbrain. Limb bud labeling began at stage 20, whereas expression in the branchial arches appeared at stage 25. Chick DAN expression generally corresponded to that of mouse DAN expression as shown by comparative in situ hybridization. However, chick DAN was found in the otic epithelium and notochord, whereas mouse DAN was restricted to the overlying otic ectomesenchyme and was absent from the notochord. This observation suggests that DAN may play different roles in chick and mouse otic and notochord development. (C) 2002 Wiley-Liss, Inc.