Soto Julio G, White Stephanie A, Reyes Steven R, Regalado Renato, Sanchez Elda E, Perez John C
Biological Sciences Department, San Jose State University, One Washington Square, Duncan Hall 254, San Jose, CA 95192-0100, United States.
Gene. 2007 Mar 1;389(1):66-72. doi: 10.1016/j.gene.2006.09.020. Epub 2006 Oct 5.
Several types of disintegrins have been isolated from Crotalus spp rattlesnakes, including RGD disintegrins, and PIII-SVMPs. We isolated six cDNAs from snake venom glands using RT-PCR. Three RGD disintegrins (atroxatin, mojastin, and viridistatin) and three PIII-SVMPs (catroriarin, scutiarin, and viristiarin) cDNAs were isolated from the rattlesnakes Crotalus atrox, Crotalus scutulatus scutulatus, and Crotalus viridis viridis, respectively. Atroxatin and Viridistatin shared 90% amino acid identity to each other, and 87% identity to Mojastin. Scutiarin and Viristiarin were identical. All PIII-SVMPs isolated in this study shared the highest amino acid identity with Catrocollastatin. cDNA and protein sequences for RGD disintegrins, one MVD disintegrin, and PIII-SVMPs of the genus Crotalus (present in the NCBI database), were used in phylogenetic analysis. Neighbor-joining analysis of PIII-SVMP and RGD/MVD disintegrin-coding DNA sequences showed that these groups of genes separate into separate clades. A Phi(ST) pairwise comparison and Analysis of Molecular Variance (AMOVA) between PIII-SVMPs and RGD/MVD disintegrins showed significant genetic differences. Mutations observed in ten of the cDNAs analyzed did not affect Cys-coding sequences. Our K(A)/K(S) data suggest that rapid evolution occurred between the genes coding for PIII-SVMPs resulting, in the production of RGD disintegrin-coding genes. However, once these genes diverged, mutations in the PIII-SVMP-coding genes were accumulated less frequently.
已从响尾蛇属的响尾蛇中分离出几种类型的去整合素，包括RGD去整合素和PIII-SVMPs。我们使用逆转录聚合酶链反应（RT-PCR）从蛇毒腺中分离出六个互补DNA（cDNA）。分别从墨西哥矛头蝮、斑纹响尾蛇指名亚种和草原响尾蛇中分离出三个RGD去整合素（atroxatin、mojastin和viridistatin）的cDNA以及三个PIII-SVMPs（catroriarin、scutiarin和viristiarin）的cDNA。Atroxatin和Viridistatin彼此的氨基酸同一性为90%，与Mojastin的同一性为87%。Scutiarin和Viristiarin是相同的。本研究中分离出的所有PIII-SVMPs与Catrocollastatin的氨基酸同一性最高。响尾蛇属的RGD去整合素、一种MVD去整合素和PIII-SVMPs的cDNA及蛋白质序列（存在于NCBI数据库中）用于系统发育分析。对PIII-SVMP和RGD/MVD去整合素编码DNA序列进行邻接法分析表明，这些基因组分成不同的进化枝。PIII-SVMPs与RGD/MVD去整合素之间的Phi(ST)成对比较和分子方差分析（AMOVA）显示出显著的遗传差异。在所分析十个cDNA中观察到的突变不影响半胱氨酸编码序列。我们的Ka/Ks数据表明，编码PIII-SVMPs的基因之间发生了快速进化，从而产生了编码RGD去整合素的基因。然而，一旦这些基因发生分歧，PIII-SVMP编码基因中的突变积累频率就较低。
