Guo X D, Kramer J M
Laboratory for Cell, Molecular, and Developmental Biology, University of Illinois, Chicago 60680.
J Biol Chem. 1989 Oct 15;264(29):17574-82.
We have identified and characterized the two genes, clb-1 and clb-2, that encode basement membrane collagen (IV) in Caenorhabditis elegans. Both genes encode 5.5-kilobase mRNAs, similar in size to the mammalian and Drosophila type IV collagen gene transcripts but much larger than the cuticle collagen transcripts of C. elegans. The nucleotide sequences of the NC1 regions of both genes were determined. Comparisons of the clb-1 and clb-2 NC1 amino acid sequences with those of mouse and human show that clb-1 shares 72% identity with mammalian alpha 2(IV) and clb-2 shares 63% identity with mammalian alpha 1(IV), suggesting that clb-1 is the alpha 2(IV) homologue and clb-2 is the alpha 1(IV) homologue. The presence of two type IV collagen genes in C. elegans and the mammals, but a single gene in Drosophila, indicates that the primordial type IV collagen gene had already duplicated early in the evolution of the invertebrates and that one of the genes has been subsequently lost from Drosophila. The mouse and human alpha 1(IV) and alpha 2(IV) genes are separated by only about 130 base pairs and are transcribed in opposite directions from overlapping promoters. We show that the C. elegans genes are located on separate chromosomes, clb-1 on X and clb-2 on III, demonstrating that the mammalian arrangement is not a requirement for all type IV collagen genes. We have identified a candidate genetic locus for the clb-2 basement membrane collagen gene, which will allow us to pursue a genetic analysis of basement membrane structure and function in C. elegans.
我们已经鉴定并表征了秀丽隐杆线虫中编码基底膜胶原蛋白(IV型)的两个基因,即clb-1和clb-2。这两个基因都编码5.5千碱基的mRNA,其大小与哺乳动物和果蝇的IV型胶原蛋白基因转录本相似,但比秀丽隐杆线虫的表皮胶原蛋白转录本大得多。测定了这两个基因NC1区域的核苷酸序列。将clb-1和clb-2的NC1氨基酸序列与小鼠和人类的序列进行比较,结果表明clb-1与哺乳动物的α2(IV)有72%的同一性,clb-2与哺乳动物的α1(IV)有63%的同一性,这表明clb-1是α2(IV)的同源物,clb-2是α1(IV)的同源物。秀丽隐杆线虫和哺乳动物中存在两个IV型胶原蛋白基因,而果蝇中只有一个基因,这表明原始的IV型胶原蛋白基因在无脊椎动物进化早期就已经复制,并且其中一个基因随后在果蝇中丢失。小鼠和人类的α1(IV)和α2(IV)基因仅相隔约130个碱基对,并从重叠的启动子以相反的方向转录。我们发现秀丽隐杆线虫的这两个基因位于不同的染色体上,clb-1在X染色体上,clb-2在III号染色体上,这表明哺乳动物的这种排列方式并非所有IV型胶原蛋白基因所必需。我们已经确定了clb-2基底膜胶原蛋白基因的一个候选遗传位点,这将使我们能够对线虫基底膜的结构和功能进行遗传分析。