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海胆的Spec 1样基因的转录会因细胞外基质的破坏而受到选择性抑制。

Transcription of the Spec 1-like gene of Lytechinus is selectively inhibited in response to disruption of the extracellular matrix.

作者信息

Wessel G M, Zhang W, Tomlinson C R, Lennarz W J, Klein W H

机构信息

Department of Biochemistry and Molecular Biology, University of Texas M.D. Anderson Cancer Center, Houston 77030.

出版信息

Development. 1989 Jun;106(2):355-65. doi: 10.1242/dev.106.2.355.

DOI:10.1242/dev.106.2.355
PMID:2591320
Abstract

The influence of the extracellular matrix (ECM) on differential gene expression during sea urchin development was explored using cell-type-specific cDNA probes. The ECM of three species of sea urchin, Strongylocentrotus purpuratus, Lytechinus variegatus and Lytechinus pictus, was disrupted with the lathrytic agent beta-aminopropionitrile (BAPN), which inhibits collagen deposition in the ECM and arrests gastrulation (Wessel & McClay, Devl Biol. 121: 149, 1987). The levels of several mRNAs (Spec 1, Spec 2, CyIIa actin, CyIIIa actin and collagen in S. purpuratus, and metallothionine, ubiquitin and LpS3 in L. pictus and L. variegatus) were compared in BAPN-treated and control embryos. These mRNAs accumulated normally during BAPN treatment, even though the embryos did not gastrulate. To determine if the expression of any gene product is sensitive to ECM disruption, a differential cDNA screen compared poly (A+) RNA from BAPN-arrested and control embryos in Lytechinus. A cDNA clone was isolated from this screen that represented a 2.1 kb mRNA that did not accumulate during BAPN treatment. Removal of BAPN resulted in the accumulation of this transcript coincident with the onset of gastrulation. This cDNA clone encodes a L. variegatus homologue of LpS1, recently demonstrated to be an ancestral homologue of the aboral ectoderm-specific Spec 1-Spec 2 gene family in S. purpuratus. Nuclear run-on assays in L. pictus suggested that transcriptional activity of LpS1 was selectively inhibited by BAPN treatment. Thus, although the accumulation of many gene products occurred independently of the embryonic collagenous matrix, the accumulation of LpS1 and LvS1 appeared to be mediated by the ECM.

摘要

利用细胞类型特异性cDNA探针,研究了细胞外基质(ECM)对海胆发育过程中差异基因表达的影响。用溶菌剂β-氨基丙腈(BAPN)破坏了三种海胆(紫球海胆、多斑刺海胆和花斑刺海胆)的ECM,该试剂可抑制ECM中胶原蛋白的沉积并阻止原肠胚形成(韦塞尔和麦克莱,《发育生物学》。121:149,1987)。比较了BAPN处理组和对照组胚胎中几种mRNA(紫球海胆中的Spec 1、Spec 2、CyIIa肌动蛋白、CyIIIa肌动蛋白和胶原蛋白,以及多斑刺海胆和花斑刺海胆中的金属硫蛋白、泛素和LpS3)的水平。这些mRNA在BAPN处理期间正常积累,尽管胚胎没有进行原肠胚形成。为了确定是否有任何基因产物的表达对ECM破坏敏感,进行了差异cDNA筛选,比较了多斑刺海胆中BAPN阻滞胚胎和对照胚胎的聚腺苷酸(A+)RNA。从该筛选中分离出一个cDNA克隆,它代表一个2.1 kb的mRNA,在BAPN处理期间不积累。去除BAPN后,该转录本的积累与原肠胚形成的开始同时发生。该cDNA克隆编码多斑刺海胆LpS1的同源物,最近已证明它是紫球海胆口外胚层特异性Spec 1-Spec 2基因家族的祖先同源物。多斑刺海胆的核延伸分析表明,BAPN处理选择性抑制了LpS1的转录活性。因此,尽管许多基因产物的积累独立于胚胎胶原基质发生,但LpS1和LvS1的积累似乎是由ECM介导的。

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