Nchongboh Chofong Gilbert, Wu Guan-Wei, Hong Ni, Wang Guo-Ping
National Key Laboratory of Agromicrobiology, Huazhong Agricultural University, Wuhan, 430070, Hubei, People's Republic of China.
Virus Genes. 2014 Dec;49(3):456-65. doi: 10.1007/s11262-014-1100-x. Epub 2014 Jul 27.
Citrus tristeza virus (CTV) is one of the most devastating pathogens of citrus. Its genome is organized into 12 open reading frames (ORFs), of which ten ORFs located at the 3'-terminus of the genome have multiple biological functions. The ten genes at the 3'-terminus of the genome of a severe isolate (CTV-S4) and three ORFs (CP, CPm and p20) of three other isolates (N4, S45 and HB1) were cloned into pGBKT7 and pGADT7 yeast shuttle vectors. Yeast two-hybridization (Y2H) assays results revealed a strong self-interaction for CP and p20, and a unique interaction between the CPm of CTV-S4 (severe) and CP of CTV-N4 (mild) isolates. Bimolecular fluorescence complementation also confirmed these interactions. Analysis of the deletion mutants delineated the domains of CP and p20 self-interaction. Furthermore, the domains responsible for CP and p20 self-interactions were mapped at the CP amino acids sites 41-84 and p20 amino acids sites 1-21 by Y2H. This study provided new information on CTV protein interactions which will help for further understanding the biological functions.
柑橘衰退病毒（CTV）是柑橘最具毁灭性的病原体之一。其基因组由12个开放阅读框（ORF）组成，其中位于基因组3'末端的10个ORF具有多种生物学功能。将一个强毒株（CTV-S4）基因组3'末端的10个基因以及其他三个毒株（N4、S45和HB1）的三个ORF（CP、CPm和p20）克隆到pGBKT7和pGADT7酵母穿梭载体中。酵母双杂交（Y2H）分析结果显示，CP和p20存在强烈的自身相互作用，且CTV-S4（强毒株）的CPm与CTV-N4（弱毒株）的CP之间存在独特的相互作用。双分子荧光互补也证实了这些相互作用。对缺失突变体的分析确定了CP和p20自身相互作用的结构域。此外，通过Y2H将负责CP和p20自身相互作用的结构域定位在CP的第41-84位氨基酸位点和p20的第1-21位氨基酸位点。本研究提供了关于CTV蛋白相互作用的新信息，这将有助于进一步了解其生物学功能。
