de la Fuente José, Maritz-Olivier Christine, Naranjo Victoria, Ayoubi Patricia, Nijhof Ard M, Almazán Consuelo, Canales Mario, Pérez de la Lastra José M, Galindo Ruth C, Blouin Edmour F, Gortazar Christian, Jongejan Frans, Kocan Katherine M
Department of Veterinary Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK 74078, USA.
BMC Genomics. 2008 Aug 2;9:372. doi: 10.1186/1471-2164-9-372.
Subolesin is an evolutionary conserved protein that was discovered recently in Ixodes scapularis as a tick protective antigen and has a role in tick blood digestion, reproduction and development. In other organisms, subolesin orthologs may be involved in the control of developmental processes. Because of the profound effect of subolesin knockdown in ticks and other organisms, we hypothesized that subolesin plays a role in gene expression, and therefore affects multiple cellular processes. The objective of this study was to provide evidence for the role of subolesin in gene expression.
Two subolesin-interacting proteins were identified and characterized by yeast two-hybrid screen, co-affinity purification and RNA interference (RNAi). The effect of subolesin knockdown on the tick gene expression pattern was characterized by microarray analysis and demonstrated that subolesin RNAi affects the expression of genes involved in multiple cellular pathways. The analysis of subolesin and interacting protein sequences identified regulatory motifs and predicted the presence of conserved protein kinase C (PKC) phosphorylation sites.
Collectively, these results provide evidence that subolesin plays a role in gene expression in ticks.
亚油酸结合蛋白是一种进化保守的蛋白质,最近在肩突硬蜱中作为蜱保护性抗原被发现,并且在蜱的血液消化、繁殖和发育中起作用。在其他生物体中,亚油酸结合蛋白直系同源物可能参与发育过程的调控。由于亚油酸结合蛋白敲低在蜱和其他生物体中具有深远影响,我们推测亚油酸结合蛋白在基因表达中起作用,因此影响多个细胞过程。本研究的目的是为亚油酸结合蛋白在基因表达中的作用提供证据。
通过酵母双杂交筛选、共亲和纯化和RNA干扰(RNAi)鉴定并表征了两种与亚油酸结合蛋白相互作用的蛋白质。通过微阵列分析表征了亚油酸结合蛋白敲低对蜱基因表达模式的影响,结果表明亚油酸结合蛋白RNA干扰影响参与多个细胞途径的基因表达。对亚油酸结合蛋白和相互作用蛋白序列的分析鉴定了调控基序,并预测了保守的蛋白激酶C(PKC)磷酸化位点的存在。
总体而言,这些结果提供了证据,证明亚油酸结合蛋白在蜱的基因表达中起作用。