Local and distant sequences are required for efficient readthrough of the barley yellow dwarf virus PAV coat protein gene stop codon
Local and distant sequences are required for efficient readthrough of the barley yellow dwarf virus PAV coat protein gene stop codon
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DOI：
10.1128/jvi.70.9.5884-5892.1996
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发表时间：
1996-09-01
影响因子：
5.4
通讯作者：
Miller, WA
中科院分区：
文献类型：
--
作者：
Brown, CM;DineshKumar, SP;Miller, WA
许多病毒使用终止密码子通读作为产生延伸外壳或复制酶蛋白的策略。大麦黄矮病毒（PAV血清型）外壳蛋白基因的终止密码子以低速率通读。这产生了一个延伸的多肽，成为病毒体的一部分。我们已经分析了大麦黄矮病毒PAV基因组中的顺式作用序列所需的程序通读在体外小麦胚芽提取物和网织红细胞裂解物和在体内燕麦原生质体。需要终止密码子3'的两个区域。删除位于终止密码子3'端的16个CCN NNN重复序列中的前5个片段可大大降低体外和体内的通读。令人惊讶的是，通读还需要位于通读开放阅读框内终止密码子3'的697至758个碱基处的第二个更远端的元件。该元件也在体内燕麦原生质体中的功能时，放置超过2 kb的外壳蛋白基因停止在非翻译区后的GUS报告基因。这是第一次报道病毒中的长距离通读信号。
Many viruses use stop codon readthrough as a strategy to produce extended coat or replicase proteins. The stop codon of the barley yellow dwarf virus (PAV serotype) coat protein gene is read through at a low rate. This produces an extended polypeptide which becomes part of the virion. We have analyzed the cis-acting sequences in the barley yellow dwarf virus PAV genome required for this programmed readthrough in vitro in wheat germ extracts and reticulocyte lysates and in vivo in oat protoplasts. Two regions 3' to the stop codon were required. Deletion of sections containing the first 5 of the 16 CCN NNN repeats located 3' of the stop codon greatly reduced readthrough in vitro and in vivo. Surprisingly, readthrough also required a second, more distal element that is located 697 to 758 bases 3' of the stop codon within the readthrough open reading frame. This element also functioned in vivo in oat protoplasts when placed more than 2 kb from the coat protein gene stop in the untranslated region following a GUS reporter gene. This is the first report of a long-range readthrough signal in viruses.