Tan Angelina W L, Francischetti Ivo M B, Slovak Mirko, Kini R Manjunatha, Ribeiro José M C
NUS Graduate School for Integrative Sciences and Engineering, National University of Singapore, Singapore 117456, Singapore; Department of Biological Sciences, Faculty of Science, National University of Singapore, Singapore 117543, Singapore.
Vector Biology Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville MD 20852, USA.
J Proteomics. 2015 Mar 18;117:120-44. doi: 10.1016/j.jprot.2014.12.014. Epub 2015 Jan 7.
Ticks rely exclusively on vertebrate blood for their survival. During feeding ticks inject into their hosts a sophisticated salivary potion that overcomes host hemostasis and adverse inflammatory responses. These mediators may also enhance pathogen transmission. Knowledge of the tick salivary protein repertoire may lead to vaccine targets to disrupt feeding and/or parasite transmission as well as to the discovery of novel pharmacological agents. Male saliva may also assist reproduction because males use their mouthparts to lubricate and introduce their spermatophores into the females' genital pore. The analyses of the sialomes of male and female ticks independently allow us to understand the strategy used by each gender to feed successfully. We sequenced cDNA libraries from pools of salivary glands from adult male and female Rhipicephalus pulchellus feeding at different time points, using the Illumina HiSeq protocol. De novo assembly of a total of 241,229,128 paired-end reads lead to extraction of 50,460 coding sequences (CDS), 11,277 of which had more than 75% coverage to known transcripts, or represented novel sequences, and were submitted to GenBank. Additionally, we generated the proteome, from the salivary gland extracts of male and female R. pulchellus, yielding a total of 454 and 2063 proteins respectively which were identified by one or more peptides with at least 95% confidence. The data set is presented as an annotated hyperlinked Excel spreadsheet, describing 121 putative secreted protein families. Female and male specific transcripts were identified.
This annotated R. pulchellus database represents a mining field for future experiments involving the resolution of time-dependent transcript expression in this tick species, as well as to define novel vaccine targets and discover novel pharmaceuticals. Gender specific proteins may represent different repertoires of pharmacological reagents to assist feeding by each sex, and in males may represent proteins that assist reproduction similarly to seminal proteins in other animals.
蜱虫完全依赖脊椎动物血液生存。在进食过程中,蜱虫会向宿主注入一种复杂的唾液混合物,以克服宿主的止血和不良炎症反应。这些介质也可能增强病原体传播。了解蜱虫唾液蛋白库可能会带来破坏进食和/或寄生虫传播的疫苗靶点,以及发现新型药物。雄性唾液也可能有助于繁殖,因为雄性用口器润滑并将精荚引入雌性生殖孔。分别分析雄性和雌性蜱虫的唾液腺蛋白质组,使我们能够了解每种性别成功进食所采用的策略。我们使用Illumina HiSeq协议对成年雄性和雌性微小扇头蜱在不同时间点进食的唾液腺池的cDNA文库进行了测序。对总共241,229,128对末端读数进行从头组装,提取出50,460个编码序列(CDS),其中11,277个与已知转录本的覆盖率超过75%,或代表新序列,并提交给了GenBank。此外,我们从雄性和雌性微小扇头蜱的唾液腺提取物中生成了蛋白质组,分别产生了总共454种和2063种蛋白质,这些蛋白质由一种或多种肽以至少95%的置信度鉴定。数据集以带注释的超链接Excel电子表格形式呈现,描述了121个假定的分泌蛋白家族。鉴定出了雌性和雄性特异性转录本。
这个带注释的微小扇头蜱数据库代表了一个挖掘领域,可用于未来涉及解析该蜱虫物种中时间依赖性转录本表达的实验,以及定义新的疫苗靶点和发现新型药物。性别特异性蛋白质可能代表不同的药理试剂库,以协助每种性别的进食,在雄性中可能代表类似于其他动物精液蛋白的有助于繁殖的蛋白质。