Junqueira-de-Azevedo Inácio de L M, Ho Paulo L
Centro de Biotecnologia, Instituto Butantan, Av. Vital Brazil, 1500, São Paulo, SP 05503-900, Brazil.
Gene. 2002 Oct 16;299(1-2):279-91. doi: 10.1016/s0378-1119(02)01080-6.
In order to produce a global panorama of the transcriptional activity of snake venom glands and to correlate with its venom composition, we constructed a DNA complementary to RNA library from the venom glands of the Viperidae snake Bothrops insularis for the generation of expressed sequence tags (ESTs). Sequences from 610 independent clones were grouped in 297 clusters, revealing the putative identification of 210 distinct gene products. Toxin sequences correspond to 56% of all transcripts (85 clusters), being the metalloproteinases (23%) and the bradykinin-potentiating peptides (11%) the major components. This approach revealed a new highly expressed toxin similar to vascular endothelial growth factor, which was recently reported (J. Biol. Chem. 276 (2001) 39836). Among the 125 clusters matching cellular proteins, the major part represents molecules involved in gene and protein expression, notably in disulfide bond assembly, reflecting a high specialization of this tissue for toxin synthesis. An unusual representation of retrotransposon-like sequences was also found and could be related to the occurrence and diversity of many paralogous forms of toxins in the venom gland. Our B. insularis dbEST allowed the identification of the most common classes of toxins present in Viperidae venoms, which parallels the complex hemorrhagic effects evoked by the venom on the prey. In addition, it provides the first comprehensive set of reptilian gene sequences described so far.
为了呈现蛇毒腺转录活性的全景并将其与毒液成分相关联,我们构建了一个来自岛蚺蝰蛇(Bothrops insularis)毒腺的RNA互补DNA文库,用于生成表达序列标签(ESTs)。来自610个独立克隆的序列被归为297个簇,揭示了210种不同基因产物的假定鉴定。毒素序列占所有转录本的56%(85个簇),其中金属蛋白酶(23%)和缓激肽增强肽(11%)是主要成分。这种方法揭示了一种新的高表达毒素,类似于血管内皮生长因子,最近有相关报道(《生物化学杂志》276 (2001) 39836)。在与细胞蛋白匹配的125个簇中,大部分代表参与基因和蛋白表达的分子,特别是在二硫键组装方面,这反映了该组织在毒素合成方面的高度专业化。还发现了逆转座子样序列的异常表现,这可能与毒腺中许多毒素旁系同源形式的出现和多样性有关。我们的岛蚺蝰蛇dbEST允许鉴定蝰蛇科毒液中最常见的毒素类别,这与毒液对猎物引起的复杂出血效应相似。此外,它提供了迄今为止描述的第一套完整的爬行动物基因序列。