Nonaka M, Takahashi M, Sasaki M
Department of Biochemistry, Nagoya City University Medical School, Japan.
J Immunol. 1994 Mar 1;152(5):2263-9.
To elucidate the origin and evolution of the complement system and the MHC, we isolated cDNA clones for the MHC class III complement factor B (Bf) gene from lamprey, one of the most primitive extant vertebrates. A part of the serine protease domain of the lamprey Bf was amplified by reverse transcriptase-PCR using the degenerated primers corresponding to the conserved amino acid stretches between the mouse Bf and C2 sequences. A full-length lamprey Bf cDNA clone was isolated from the lamprey liver cDNA library using a PCR-amplified DNA clone as a probe. The deduced amino acid sequence of 763 residues showed essentially the same domain structure as mammalian Bf or C2, consisting of three short consensus repeat domains, a von Willebrand domain, and a serine protease domain. Lamprey Bf showed 33 and 29% overall amino acid similarity to mouse Bf and mouse C2, respectively, whereas amino acid similarity between mouse Bf and mouse C2 was 36%, suggesting that the gene duplication of Bf/C2 occurred in the main line of vertebrate evolution after the divergence of cyclostomes. This is the first report of the molecular cloning from cyclostomes of a component of the mammalian MHC that offers the possibility of genetic analysis of the presumably primitive MHC of cyclostomes.
为阐明补体系统和主要组织相容性复合体(MHC)的起源与进化,我们从现存最原始的脊椎动物之一七鳃鳗中分离出了MHC III类补体因子B(Bf)基因的cDNA克隆。使用与小鼠Bf和C2序列之间保守氨基酸序列相对应的简并引物,通过逆转录聚合酶链反应(RT-PCR)扩增了七鳃鳗Bf的丝氨酸蛋白酶结构域的一部分。以PCR扩增的DNA克隆为探针,从七鳃鳗肝脏cDNA文库中分离出全长七鳃鳗Bf cDNA克隆。推导的763个残基的氨基酸序列显示出与哺乳动物Bf或C2基本相同的结构域结构,由三个短共有重复结构域、一个血管性血友病因子结构域和一个丝氨酸蛋白酶结构域组成。七鳃鳗Bf与小鼠Bf和小鼠C2的总体氨基酸相似性分别为33%和29%,而小鼠Bf和小鼠C2之间的氨基酸相似性为36%,这表明Bf/C2的基因复制发生在圆口纲动物分化后的脊椎动物进化主线中。这是首次从圆口纲动物中克隆出哺乳动物MHC成分的分子,这为对推测的原始圆口纲动物MHC进行遗传分析提供了可能性。