Craniofacial dysmorphogenesis including cleft palate in mice with an insertional mutation in the discs large gene
Craniofacial dysmorphogenesis including cleft palate in mice with an insertional mutation in the discs large gene
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DOI：
10.1128/mcb.21.5.1475-1483.2001
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发表时间：
2001-03-01
影响因子：
5.3
通讯作者：
Bernstein, A
中科院分区：
文献类型：
--
作者：
Caruana, G;Bernstein, A
椎间盘大 (DIg) 蛋白或突触相关蛋白 97 (SAP97) 是多域支架蛋白膜相关鸟苷酸激酶家族的成员，该蛋白将跨膜和信号分子募集到局部质膜位点。小鼠 dig 是果蝇 dig 肿瘤抑制基因的同源物。果蝇dig功能的丧失会破坏成虫盘上皮细胞的细胞生长控制、顶端基底极性和细胞粘附，导致胚胎致死。在这项研究中，我们通过全能胚胎干细胞中的基因捕获分离了小鼠 dig 基因座中的突变插入。该插入导致截短的蛋白质产物，其中包含与 LacZ 报告基因融合的 Dig 的 N 端三个 PSD-95/DLG/ZO-1 结构域，随后缺少 src 同源 3 (SH3)、蛋白质 4.1 结合和鸟苷酸激酶 (GUK) 样结构域。 Dlg-LacZ 融合蛋白在胚胎发生过程中在上皮细胞、间充质细胞、神经元细胞、内皮细胞和造血细胞中表达。 dig 突变纯合子小鼠在子宫内表现出生长迟缓，前颌骨和下颌骨发育不全？患有第二腭裂，并在围产期死亡。与这种表型一致，Dlg-LacZ 在整个腭发育过程中在间充质和上皮细胞中表达。我们对 dlg 突变小鼠的遗传和表型分析表明，涉及 SH3、蛋白质 4.1 结合和/或 GUK 样结构域的蛋白质-蛋白质相互作用对于小鼠 Dig 在颅面和腭形态发生中的正常功能至关重要。
The discs large (DIg) protein, or synapse-associated protein 97 (SAP97), is a member of the membrane-associated guanylate kinase family of multidomain scaffolding proteins which recruits transmembrane and signaling molecules to localized plasma membrane sites. Murine dig is the homologue of the Drosophila dig tumor suppressor gene. The loss of dig function in Drosophila disrupts cellular growth control, apicobasal polarity, and cell adhesion of imaginal disc epithelial cells, resulting in embryonic lethality. In this study, we isolated a mutational insertion in the murine dig locus by gene trapping in totipotent embryonic stem cells. This insertion results in a truncated protein product that contains the N-terminal three PSD-95/DLG/ZO-1 domains of Dig fused to the LacZ reporter and subsequently lacks the src homology 3 (SH3), protein 4.1 binding, and guanylate kinase (GUK)-like domains. The Dlg-LacZ fusion protein is expressed in epithelial, mesenchymal, neuronal, endothelial, and hematopoietic cells during embryogenesis. Mice homozygous for the dig mutation exhibit growth retardation in utero, have hypoplasia of the premaxilla and mandible? have a cleft secondary palate, and die perinatally. Consistent with this phenotype, Dlg-LacZ is expressed in mesenchymal and epithelial cells throughout palatal development. Our genetic and phenotypic analysis of dlg mutant mice suggests that protein-protein interactions involving the SH3, protein 4.1 binding, and/or GUK-like domains are essential to the normal function of murine Dig within craniofacial and palatal morphogenesis.