Functional analysis of protein disulfide isomerases in blood feeding, viability and oocyte development in Haemaphysalis longicornis ticks
Functional analysis of protein disulfide isomerases in blood feeding, viability and oocyte development in Haemaphysalis longicornis ticks
复制标题
DOI：
10.1016/j.ibmb.2007.11.006
复制
发表时间：
2008-03-01
影响因子：
3.8
通讯作者：
Fujisaki, Kozo
中科院分区：
文献类型：
--
作者：
Liao, Min;Boldbaatar, Daindinsuren;Fujisaki, Kozo
先前从长角血蜱蜱（指定为H1PDI-1， H1PDI-2和H1PDI-3）中鉴定出三种蛋白质二硫异构酶。为了进一步分析它们的生物学功能，我们将每个H1PDI基因的dsRNA和H1PDI-1/H1PDI-3的一个dsRNA组合分别注射到雌性蜱体内。real-time PCR、RT-PCR和Western blot分析证实RNA干扰（RNAi）可降低H1PDIs基因和蛋白的表达。在单注射dsrna组中，H1PDI-1 RNAi影响蜱的吸血和产卵，H1PDI-2 RNAi影响蜱的生存能力，而H1PDI-3 RNAi本身无显著影响。然而，联合注射H1PDI-1/H1PDI-3 dsRNA对蜱的生存能力有协同作用。此外，注射了H1PDI-2 dsrna的蜱和注射了H1PDI-1/H1PDI-3 dsrna的蜱分别严重破坏了中肠和表皮，并且H1PDI基因的破坏导致蜱中含二硫键的卵黄原蛋白（Vg）表达显著降低。这些结果表明，长角蜱的pdi可能通过介导蜱体内含二硫键蛋白的形成以及基膜和角质层成分如细胞外基质（ECM）的形成，参与了蜱的供血、生存和卵母细胞发育。本文首次报道了PDI家族分子在血食性节肢动物中的功能分析以及与其他分子的相互作用。(C) 2007 Elsevier Ltd.版权所有。
Three protein disulfide isomerases from Haemaphysalis longicornis ticks (designated as H1PDI-1, H1PDI-2, and H1PDI-3) were previously identified. In order to further analyze their biological functions, the dsRNA of each H1PDI gene and one dsRNA combination of H1PDI-1/H1PDI-3 were separately injected into female ticks. Reduction of gene and protein expression of H1PDIs by RNA interference (RNAi) was demonstrated by real-time PCR, RT-PCR and Western blot analysis. In single dsRNA-injected groups, H1PDI-1 RNAi impacted tick blood feeding and oviposition, H1PDI-2 RNAi impacted tick viability and H1PDI-3 RNAi had no significant impact by itself. However, the injection of a combination of H1PDI-1/H1PDI-3 dsRNA had synergistic effects on tick viability. Furthermore, the midgut and cuticle were severely damaged in H1PDI-2 dsRNA-injected ticks and H1PDI-1/H1PDI-3 dsRNA-injected ticks, respectively, and disruption of H1PDI genes led to a significant reduction of disulfide bond-containing vitellogenin (Vg) expression in ticks. These results indicate that PDIs from H. longicornis are involved in blood feeding, viability and oocyte development, probably by mediating the formation of disulfide bond-containing proteins of the ticks and the formation of basement membrane and cuticle components such as extracellular matrix (ECM). This is the first report on the functional analysis of PDI family molecules as well as the interactions of PDI and other molecules in blood-feeding arthropods. (C) 2007 Elsevier Ltd. All rights reserved.