Chen Y, Faraco J, Yin W, Germiller J, Francke U, Bonadio J
Department of Pathology, University of Michigan, Ann Arbor 48109-0650.
J Biol Chem. 1993 Dec 25;268(36):27381-9.
The microfibril-associated glycoprotein (MAGP) was recently established as a discrete constituent of 10-nm microfibrils. We have characterized the primary structure of the mouse transcript, the structure and chromosomal localization of the murine gene, and the developmental pattern of gene expression. The transcript consists of 1,037 base pairs as determined by cDNA cloning, Northern blot analysis, S1 nuclease mapping, and primer extension mapping. Using a cDNA fragment as a probe, we isolated a single genomic clone that contained the entire mouse gene. Analysis of this clone indicated that Magp is fragmented into 9 exons, with the initiator Met codon located in exon 2. As determined by analysis of somatic cell hybrid lines and by fluorescence in situ hybridization, the mouse gene was mapped to chromosome 4 at a location corresponding to region D3-E1. Genomic sequence immediately upstream of the transcription start site was found to be GC-rich but lacked TATA or CCAAT boxes as well as other cis-acting motifs known to regulate transcription. Promoters of this type are usually found in genes that exhibit broad temporal and spatial patterns of expression. Consistent with this idea, the Magp transcript appeared to be the widespread product of mesenchymal/connective tissue cells throughout mouse development. This study presents the first comprehensive evaluation of microfibril gene expression during mammalian development.
微原纤维相关糖蛋白(MAGP)最近被确定为10纳米微原纤维的一种离散成分。我们已经对小鼠转录本的一级结构、小鼠基因的结构和染色体定位以及基因表达的发育模式进行了表征。通过cDNA克隆、Northern印迹分析、S1核酸酶图谱分析和引物延伸图谱分析确定,该转录本由1037个碱基对组成。我们使用一个cDNA片段作为探针,分离出了一个包含整个小鼠基因的单一基因组克隆。对该克隆的分析表明,Magp被分割为9个外显子,起始甲硫氨酸密码子位于外显子2中。通过对体细胞杂交系的分析和荧光原位杂交确定,小鼠基因定位于4号染色体上对应于区域D3 - E1的位置。发现在转录起始位点上游紧邻的基因组序列富含GC,但缺乏TATA或CCAAT框以及其他已知调控转录的顺式作用基序。这种类型的启动子通常存在于表现出广泛的时间和空间表达模式的基因中。与此观点一致,Magp转录本似乎是整个小鼠发育过程中间充质/结缔组织细胞广泛产生的产物。这项研究首次对哺乳动物发育过程中的微原纤维基因表达进行了全面评估。