Saurav Gunjan Kumar, Rana Vipin Singh, Popli Sonam, Daimei Guisuibou, Rajagopal Raman
Gut Biology Laboratory, Department of Zoology, University of Delhi, Room No. 117, Delhi, 110007, India.
Dr. B. R. Ambedkar Center for Biomedical Research, University of Delhi, Delhi, 110007, India.
Virus Genes. 2019 Jun;55(3):356-367. doi: 10.1007/s11262-019-01657-z. Epub 2019 Mar 28.
Bemisia tabaci (whitefly) is the sole vector of begomoviruses, which transmits them in a persistent and circulative manner from infected to healthy plants. During this process, begomoviruses interact with various proteins in the insect vector B. tabaci that would play a specific role in the virus transmission. Identification and characterization of such proteins are important to understand the complete process of virus transmission. Coat protein (CP) of begomoviruses is the only protein which is reported to interact with proteins of the insect vector B. tabaci. In this study, we performed yeast two-hybrid assay using CP of cotton leaf curl Rajasthan virus (CLCuV) and Tomato leaf curl New Delhi virus (ToLCNDV) as bait in separate experiments and cDNA prepared from total RNA of B. tabaci was used as prey. Yeast two-hybrid assay resulted in identification of a thioredoxin-like protein (TLP) from CLCuV yeast two-hybrid library. Later TLP was also found to interact with CP of ToLCNDV. In vitro pull-down assay showed TLP interaction with CP of both CLCuV and ToLCNDV. TLP was found to interact with ToLCNDV virus particles isolated from tomato leaves.
烟粉虱是双生病毒的唯一传播媒介，它以持久循环的方式将病毒从受感染植株传播到健康植株。在此过程中，双生病毒与烟粉虱昆虫媒介中的多种蛋白质相互作用，这些蛋白质在病毒传播中发挥特定作用。鉴定和表征此类蛋白质对于了解病毒传播的完整过程很重要。双生病毒的外壳蛋白（CP）是唯一据报道与烟粉虱昆虫媒介蛋白质相互作用的蛋白质。在本研究中，我们分别使用拉贾斯坦邦棉花卷叶病毒（CLCuV）和新德里番茄卷叶病毒（ToLCNDV）的CP作为诱饵进行酵母双杂交试验，并将从烟粉虱总RNA制备的cDNA用作猎物。酵母双杂交试验从CLCuV酵母双杂交文库中鉴定出一种硫氧还蛋白样蛋白（TLP）。后来还发现TLP与ToLCNDV的CP相互作用。体外下拉试验表明TLP与CLCuV和ToLCNDV的CP均有相互作用。发现TLP与从番茄叶片中分离的ToLCNDV病毒粒子相互作用。
