Division of Gene Function and Regulation, Institute of Life Sciences, Government of India, Chandrasekharpur, Bhubaneswar, Odisha, India.
Department of Molecular Plant Virology and Plant Genetic Engineering, KTRDC, College of Agriculture, Food and Environment, University of Kentucky, Lexington, KY 40546-0236, United States.
Biochim Biophys Acta Gene Regul Mech. 2018 Feb;1861(2):133-146. doi: 10.1016/j.bbagrm.2018.01.005. Epub 2018 Feb 3.
Caulimoviral promoters have become excellent tools for efficient transgene expression in plants. However, the transcriptional framework controlling their systematic regulation is poorly understood. To understand this regulatory mechanism, we extensively studied a novel caulimoviral promoter, PV8 (-163 to +138, 301 bp), isolated from Petunia vein-clearing virus (PVCV). PVCV was found to be Salicylic acid (SA)-inducible and 2.5-3.0 times stronger than the widely used CaMV35S promoter. In silico analysis of the PV8 sequence revealed a unique clustering of two stress-responsive cis-elements, namely, as-1 and W-box, located within a span of 31 bp (-74 to -47) that bound to the TGA1a and WRKY71 plant transcription factors (TFs), respectively. We found that as-1 (TTACG) and W-box (TGAC) elements occupied both TGA1a and WRKY71 on the PV8 backbone. Mutational studies demonstrated that the combinatorial influence of as-1 (-57) and W-box (-74 and -47) on the PV8 promoter sequence largely modulated its activity. TGA1a and WRKY71 physically interacted and cooperatively enhanced the transcriptional activity of the PV8 promoter. Biotic stress stimuli induced PV8 promoter activity by ~1.5 times. We also established the possible pathogen-elicitor function of AtWRKY71 and NtabWRKY71 TFs. Altogether, this study elucidates the interplay between TFs, biotic stress and caulimoviral promoter function.
caulimoviral 启动子已成为在植物中高效表达转基因的优秀工具。然而,其系统调控的转录框架仍知之甚少。为了理解这种调控机制,我们广泛研究了一种新型 caulimoviral 启动子 PV8(-163 至+138,301bp),该启动子从 Petunia vein-clearing virus(PVCV)中分离得到。发现 PVCV 受水杨酸(SA)诱导,其活性比广泛使用的 CaMV35S 启动子强 2.5-3.0 倍。对 PV8 序列的计算机分析揭示了两个应激响应顺式元件的独特聚类,即 as-1 和 W-box,位于 31bp(-74 至-47)的跨度内,分别与 TGA1a 和 WRKY71 植物转录因子(TFs)结合。我们发现 as-1(TTACG)和 W-box(TGAC)元件占据了 PV8 骨架上的 TGA1a 和 WRKY71。突变研究表明,as-1(-57)和 W-box(-74 和-47)的组合影响极大地调节了 PV8 启动子序列的活性。TGA1a 和 WRKY71 物理相互作用并协同增强了 PV8 启动子的转录活性。生物胁迫刺激使 PV8 启动子活性增加了约 1.5 倍。我们还证实了 AtWRKY71 和 NtabWRKY71 TF 的可能病原体诱导功能。总的来说,这项研究阐明了 TF、生物胁迫和 caulimoviral 启动子功能之间的相互作用。