THE MOUSE COLLAGEN-X GENE - COMPLETE NUCLEOTIDE-SEQUENCE, EXON STRUCTURE AND EXPRESSION PATTERN
THE MOUSE COLLAGEN-X GENE - COMPLETE NUCLEOTIDE-SEQUENCE, EXON STRUCTURE AND EXPRESSION PATTERN
复制标题
DOI：
10.1042/bj2890247
复制
发表时间：
1993-01-01
影响因子：
4.1
通讯作者：
VUORIO, E
中科院分区：
文献类型：
--
作者：
ELIMA, K;EEROLA, I;VUORIO, E
从两个基因组文库中分离出覆盖7.2 kb小鼠alpha1(X)胶原蛋白基因、0.86 kb启动子和1.25 kb 3'-侧翼序列的重叠基因组克隆，并对其进行核苷酸测序。该基因的典型特征包括一个独特的三外显子结构，与鸡基因相似，整个463个氨基酸的三螺旋结构域由一个大外显子编码。在小鼠和已知的鸡、牛和人胶原X型序列之间的胶原和c端非胶原结构域的编码区，氨基酸和核苷酸序列保守程度最高。序列之间的更多分歧发生在n端非胶原结构域。哺乳动物胶原X序列之间的相似性延伸到3'-未翻译序列，特别是在聚腺苷化位点附近。小鼠胶原X基因的启动子包含两个相距159 bp的TATAA盒子；转录起始位点的引物延伸分析显示两者都是功能性的。启动子有一个不寻常的结构，在上游转录起始位点的-220和-1位置之间的G + C含量非常低，为28%。Northern和原位杂交分析证实，α 1(X)胶原基因的表达仅限于软骨内钙化组织中的肥大软骨细胞。该基因的详细序列信息可用于研究该基因的启动子活性和转基因小鼠的产生。
Overlapping genomic clones covering the 7.2 kb mouse alpha1(X) collagen gene, 0.86 kb of promoter and 1.25 kb of 3'-flanking sequences were isolated from two genomic libraries and characterized by nucleotide sequencing. Typical features of the gene include a unique three-exon structure, similar to that in the chick gene, with the entire triple-helical domain of 463 amino acids coded by a single large exon. The highest degree of amino acid and nucleotide sequence conservation was seen in the coding region for the collagenous and C-terminal non-collagenous domains between the mouse and known chick, bovine and human collagen type X sequences. More divergence between the sequences occurred in the N-terminal non-collagenous domain. Similarity between the mammalian collagen X sequences extended into the 3'-untranslated sequence, particularly near the polyadenylation site. The promoter of the mouse collagen X gene was found to contain two TATAA boxes 159 bp apart; primer extension analyses of the transcription start site revealed that both were functional. The promoter has an unusual structure with a very low G + C content of 28 % between positions -220 and -1 of the upstream transcription start site. Northern and in situ hybridization analyses confirmed that the expression of the alpha1(X) collagen gene is restricted to hypertrophic chondrocytes in tissues undergoing endochondral calcification. The detailed sequence information of the gene is useful for studies on the promoter activity of the gene and for generation of transgenic mice.