Kampkötter Andreas, Volkmann Thorsten E, de Castro Sarah Hegi, Leiers Britta, Klotz Lars Oliver, Johnson Thomas E, Link Christopher D, Henkle-Dührsen Kimberly
Institut für Genetik, Heinrich-Heine-Universitat, Universitätsstrasse 1, 40225 Dusseldorf, Germany.
J Mol Biol. 2003 Jan 3;325(1):25-37. doi: 10.1016/s0022-2836(02)01174-9.
This study examined the genomic organisation of the coding region of the glutathione S-transferase 3 (Ov-GST-3) from the human parasitic nematode Onchocerca volvulus; alternative splicing leads to three different transcripts (Ov-GST-3/1; Ov-GST-3/2 and Ov-GST-3/3). Since the expression of Ov-GST-3 is inducible by oxidative stress, it is assumed that it is involved in the defense against reactive oxygen species (ROS) resulting from cellular metabolism. Furthermore, we suggest that Ov-GST-3 plays an important role in the protection of the parasite against ROS derived from the host's immune system. To experimentally investigate these speculations, we generated Caenorhabditis elegans lines transgenic for Ov-GST-3 (AK1) and examined their resistance to artificially generated ROS. The AK1 worms (extrachromosomal and integrated lines) were found to be much more resistant to internal (juglone) and external (hypoxanthine/xanthine oxidase) oxidative stress than wild-type C.elegans worms. RNA interference experiments targeted to the Ov-GST-3 transcripts resulted in decreased resistance, confirming that this effect is due to the transgenic expression of Ov-GST-3. These results clearly demonstrate that the Ov-GST-3 gene confers an increased resistance to oxidative stress. This study also shows the applicability of C.elegans as a model organism for the functional characterization of genes from (parasitic) nematode species which are not accessible to genetic manipulations.
本研究检测了人类寄生线虫盘尾丝虫谷胱甘肽S-转移酶3(Ov-GST-3)编码区的基因组组织;可变剪接产生三种不同的转录本(Ov-GST-3/1;Ov-GST-3/2和Ov-GST-3/3)。由于Ov-GST-3的表达可被氧化应激诱导,推测它参与抵御细胞代谢产生的活性氧(ROS)。此外,我们认为Ov-GST-3在保护寄生虫免受宿主免疫系统产生的ROS侵害方面发挥重要作用。为了通过实验研究这些推测,我们构建了Ov-GST-3(AK1)转基因秀丽隐杆线虫品系,并检测它们对人工产生的ROS的抗性。发现AK1线虫(染色体外和整合品系)比野生型秀丽隐杆线虫对内部(胡桃醌)和外部(次黄嘌呤/黄嘌呤氧化酶)氧化应激的抗性要强得多。针对Ov-GST-3转录本的RNA干扰实验导致抗性降低,证实这种效应是由于Ov-GST-3的转基因表达。这些结果清楚地表明,Ov-GST-3基因赋予了对氧化应激增强的抗性。本研究还表明秀丽隐杆线虫作为一种模式生物,可用于对无法进行基因操作的(寄生)线虫物种的基因进行功能表征。