Reed Jonathan C, Kasschau Kristin D, Prokhnevsky Alexey I, Gopinath Kodetham, Pogue Gregory P, Carrington James C, Dolja Valerian V
Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331, USA.
Virology. 2003 Feb 15;306(2):203-9. doi: 10.1016/s0042-6822(02)00051-x.
Using an Agrobacterium-mediated transient assay, we screened the 15.5-kb genome of the Beet yellows virus for proteins with RNA silencing suppressor activity. Among eight proteins tested, only a 21-kDa protein (p21) was able to suppress double-stranded (ds) RNA-induced silencing of the green fluorescent protein (GFP) mRNA. Restoration of GFP expression by p21 under these conditions had no apparent effect on accumulation of the small interfering RNAs. In addition, p21 elevated the transient expression level of the GFP mRNA in the absence of dsRNA inducer. Similar activities were detected using homologs of p21 encoded by other members of the genus Closterovirus. Computer analysis indicated that p21-like proteins constitute a novel protein family that is unrelated to other recognized suppressors of RNA silencing. Examination of the subcellular distribution in BYV-infected plants revealed that p21 is partitioned between soluble cytoplasmic form and proteinaceous inclusion bodies at the cell periphery.
利用农杆菌介导的瞬时分析方法,我们在甜菜黄化病毒15.5 kb的基因组中筛选具有RNA沉默抑制活性的蛋白质。在所测试的8种蛋白质中,只有一种21 kDa的蛋白质(p21)能够抑制双链(ds)RNA诱导的绿色荧光蛋白(GFP)mRNA沉默。在此条件下,p21使GFP表达恢复,但对小干扰RNA的积累没有明显影响。此外,在没有dsRNA诱导剂的情况下,p21提高了GFP mRNA的瞬时表达水平。使用来自黄化病毒属其他成员编码的p21同源物也检测到了类似的活性。计算机分析表明,p21样蛋白构成了一个与其他已知的RNA沉默抑制子无关的新蛋白家族。对受BYV感染植物的亚细胞分布进行检测发现,p21在可溶性细胞质形式和细胞周边的蛋白质包涵体之间分配。