Sultana Hameeda, Patel Unnati, Sonenshine Daniel E, Neelakanta Girish
Center for Molecular Medicine, College of Sciences, Old Dominion University, Norfolk, 23529, VA, USA.
Department of Biological Sciences, Old Dominion University, Norfolk, 23529, VA, USA.
Parasit Vectors. 2015 Feb 28;8:132. doi: 10.1186/s13071-015-0749-x.
Subolesin is an evolutionary conserved molecule in diverse arthropod species that play an important role in the regulation of genes involved in immune responses, blood digestion, reproduction and development. In this study, we have identified a subolesin ortholog from soft ticks Ornithodoros turicata, the vector of the relapsing fever spirochete in the United States.
Uninfected fed or unfed O. turicata ticks were used throughout this study. The subolesin mRNA was amplified by reverse transcription polymerase chain reaction (RT-PCR) and sequenced. Quantitative-real time PCR (QRT-PCR) was performed to evaluate subolesin mRNA levels at different O. turicata developmental stages and from salivary glands and gut tissues. Bioinformatics and comparative analysis was performed to predict potential post-translational modifications in O. turicata subolesin amino-acid sequences.
Our study reveals that O. turicata subolesin gene expression is developmentally regulated, where; adult ticks expressed significantly higher levels in comparison to the larvae or nymphal ticks. Expression of subolesin was evident in both unfed and fed ticks and in the salivary glands and midgut tissues. The expression of subolesin transcripts varied in fed ticks with peak levels at day 14 post-feeding. Phylogenetic analysis revealed that O. turicata subolesin showed a high degree of sequence conservation with subolesin's from other soft and hard ticks. Bioinformatics and comparative analysis predicted that O. turicata subolesin carry three Protein kinase C and one Casein kinase II phosphorylation sites. However, no myristoylation or glycosylation sites were evident in the O. turicata subolesin sequence.
Our study provides important insights in recognizing subolesin as a conserved potential candidate for the development of a broad-spectrum anti-vector vaccine to control not only ticks but also several other arthropods that transmit diseases to humans and animals.
亚油酸结合蛋白是一种在多种节肢动物物种中进化保守的分子,在调节参与免疫反应、血液消化、繁殖和发育的基因方面发挥重要作用。在本研究中,我们从美国回归热螺旋体的传播媒介——软蜱土耳其钝缘蜱中鉴定出一个亚油酸结合蛋白直系同源物。
本研究全程使用未感染的饱血或未饱血的土耳其钝缘蜱。通过逆转录聚合酶链反应(RT-PCR)扩增亚油酸结合蛋白mRNA并进行测序。进行定量实时PCR(QRT-PCR)以评估土耳其钝缘蜱不同发育阶段以及唾液腺和肠道组织中亚油酸结合蛋白mRNA水平。进行生物信息学和比较分析以预测土耳其钝缘蜱亚油酸结合蛋白氨基酸序列中潜在的翻译后修饰。
我们的研究表明,土耳其钝缘蜱亚油酸结合蛋白基因表达受发育调控,其中,成年蜱的表达水平明显高于幼虫或若虫蜱。亚油酸结合蛋白在未饱血和饱血蜱以及唾液腺和中肠组织中均有明显表达。饱血蜱中亚油酸结合蛋白转录本的表达有所不同,在吸血后第14天达到峰值水平。系统发育分析表明,土耳其钝缘蜱亚油酸结合蛋白与其他软蜱和硬蜱的亚油酸结合蛋白具有高度的序列保守性。生物信息学和比较分析预测,土耳其钝缘蜱亚油酸结合蛋白有三个蛋白激酶C和一个酪蛋白激酶II磷酸化位点。然而,在土耳其钝缘蜱亚油酸结合蛋白序列中未发现肉豆蔻酰化或糖基化位点。
我们的研究为将亚油酸结合蛋白识别为开发广谱抗媒介疫苗的保守潜在候选物提供了重要见解,该疫苗不仅可以控制蜱,还可以控制其他几种将疾病传播给人类和动物的节肢动物。