An efficient ribosomal frame-shifting signal in the polymerase-encoding region of the coronavirus IBV.
An efficient ribosomal frame-shifting signal in the polymerase-encoding region of the coronavirus IBV.
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DOI：
10.1002/j.1460-2075.1987.tb02713.x
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发表时间：
1987-12-01
期刊：
影响因子：
11.4
通讯作者：
Inglis, S C
中科院分区：
文献类型：
--
作者：
Brierley, I;Boursnell, M E;Binns, M M;Bilimoria, B;Blok, V C;Brown, T D;Inglis, S C
冠状病毒传染性支气管炎病毒(IBV)基因组RNA的聚合酶编码区含有两个非常大的、短暂重叠的开放阅读框(ORF)F1和F2，根据序列分析，认为下游ORF F2的表达可能是通过核糖体移码来调节的。为了验证这一可能性，在标记基因中克隆了一个包含F1/F2重叠区的cDNA片段，并将其置于重组质粒中噬菌体SP6启动子的控制下。从该质粒转录的信使RNA，当在无细胞系统中翻译时，指定了多肽的合成，其大小与重叠区域内有效的核糖体移框事件预测的产物完全一致。产物的性质通过它们与针对侧翼标记基因的特定部分而产生的抗血清的反应来证实。这是高等真核生物中第一个核糖体框架移位的非逆转录病毒例子。
The polymerase-encoding region of the genomic RNA of the coronavirus infectious bronchitis virus (IBV) contains two very large, briefly overlapping open reading frames (ORF), F1 and F2, and it has been suggested on the basis of sequence analysis that expression of the downstream ORF, F2, might be mediated through ribosomal frame-shifting. To examine this possibility a cDNA fragment containing the F1/F2 overlap region was cloned within a marker gene and placed under the control of the bacteriophage SP6 promoter in a recombinant plasmid. Messenger RNA transcribed from this plasmid, when translated in cell-free systems, specified the synthesis of polypeptides whose size was entirely consistent with the products predicted by an efficient ribosomal frame-shifting event within the overlap region. The nature of the products was confirmed by their reactivity with antisera raised against defined portions of the flanking marker gene. This is the first non-retroviral example of ribosomal frame-shifting in higher eukaryotes.