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甜菜坏死黄脉病毒通读蛋白通读结构域的高分辨率分析:KTER基序对甜菜多黏菌高效传播该病毒至关重要。

High resolution analysis of the readthrough domain of beet necrotic yellow vein virus readthrough protein: a KTER motif is important for efficient transmission of the virus by Polymyxa betae.

作者信息

Tamada T, Schmitt C, Saito M, Guilley H, Richards K, Jonard G

机构信息

Hokkaido Central Agricultural Experiment Station, Naganuma, Japan.

出版信息

J Gen Virol. 1996 Jul;77 ( Pt 7):1359-67. doi: 10.1099/0022-1317-77-7-1359.

Abstract

The 5'-terminal cistron of beet necrotic yellow vein furovirus RNA 2 encodes the 21 kDa major viral coat protein and terminates with an amber stop codon which can undergo suppression to give rise to a 75 kDa readthrough (RT) protein referred to as P75. P75 is a minor component of virions and the 54 kDa RT domain following the coat protein sequence is important both for virus assembly and transmission by the fungal vector Polymyxa betae. To better define the regions of the RT domain involved in these two steps, RNA 2 transcripts encoding different in-frame RT domain deletion mutants were tested for their ability to form virions when inoculated to plants with the other viral RNAs and to be fungus-transmitted. All deletions in the N-terminal half of the RT domain interfered with virus assembly and partially or completely inhibited fungus transmission. A 4 1 1 nucleotide deletion within the C-terminal half of the RT domain did not inhibit assembly but blocked fungus transmission of the virus. Alanine scanning mutagenesis within the aforesaid 4 1 1 nucleotide subdomain identified a peptide motif (KTER) which is important for the fungus transmission process.

摘要

甜菜坏死黄脉真菌传棒状病毒RNA 2的5'-末端顺反子编码21 kDa的主要病毒外壳蛋白,并以琥珀色终止密码子结束,该密码子可发生抑制作用,产生一种75 kDa的通读(RT)蛋白,称为P75。P75是病毒粒子的次要成分,外壳蛋白序列之后的54 kDa RT结构域对于病毒组装和由真菌载体甜菜多黏菌传播都很重要。为了更好地确定RT结构域中参与这两个步骤的区域,对编码不同读框RT结构域缺失突变体的RNA 2转录本进行了测试,检测其与其他病毒RNA一起接种到植物时形成病毒粒子的能力以及被真菌传播的能力。RT结构域N端一半的所有缺失都干扰了病毒组装,并部分或完全抑制了真菌传播。RT结构域C端一半内的411个核苷酸缺失不抑制组装,但阻断了病毒的真菌传播。在上述411个核苷酸亚结构域内进行丙氨酸扫描诱变,确定了一个对真菌传播过程很重要的肽基序(KTER)。

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