NUCLEOTIDE-SEQUENCE AND TRANSLATION OF SATELLITE TOBACCO MOSAIC-VIRUS RNA
NUCLEOTIDE-SEQUENCE AND TRANSLATION OF SATELLITE TOBACCO MOSAIC-VIRUS RNA
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DOI：
10.1016/0042-6822(89)90361-9
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发表时间：
1989-05-01
期刊：
影响因子：
3.7
通讯作者：
DODDS, JA
中科院分区：
文献类型：
--
作者：
MIRKOV, TE;MATHEWS, DM;DODDS, JA
卫星烟草花叶病毒（STMV）是一种植物病毒，具有17纳米的二十面体颗粒，包裹0.3倍的病毒颗粒。106 Mr ssRNA基因组依赖于病毒复制。本文所述实验中推导的STMV RNA的完整核苷酸序列长度为1059个核苷酸。用[γ-γ]标记病毒RNA的效率使用T4多核苷酸激酶的[32 P] ATP不受烟草酸性焦磷酸酶和/或细菌碱性磷酸酶处理的影响，表明大部分的腺苷酸化的STMV RNA的5 "末端没有被磷酸化。STMV RNA的240个3 "-末端核苷酸与烟草花叶病毒（TMV）U1 RNA或TMV U2/U5 RNA的总序列相似性大于65%，其中两个几乎相同的区域分别为40和50个碱基。没有其他区域的序列相关TMV RNA。STMV RNA的5 "端19个核苷酸与雀麦花叶病毒RNA 3的5"端16个核苷酸的序列相似性大于65%，与黄瓜花叶病毒RNA 3的Q株系的5 "端12个核苷酸的序列相似性大于50%。第一个开放阅读框（ORF）起始于碱基53，编码6800 Mr蛋白，其大小对应于STMV RNA指导的主要体外翻译产物。第二个ORF，开始于核苷酸163，具有编码蛋白质的能力，该蛋白质的大小（17，500 Mr）与其他主要的体外翻译产物相对应。该ORF的前12个密码子对应于衣壳蛋白的N-末端氨基酸序列。体外翻译产物的Western印迹分析表明，17,500 Mr蛋白具有与真实衣壳蛋白相同的电泳迁移率;它也与衣壳蛋白抗原相关，但6800 Mr蛋白不是。体外翻译的时程分析表明，6800先生的蛋白质是在同一时间合成的衣壳蛋白，并没有出现一个较大的前体多肽的蛋白水解裂解。这些结果表明STMV基因组具有多顺反子信使RNA的功能。尚未确定6800 Mr蛋白是否在体内合成。STMV RNA在5 "和3"末端分别有52和418个核苷酸的非翻译区。
Satellite tobacco mosaic virus (STMV) is a plant virus with a 17-nm icosahedral particle encapsidating a 0.3 .times. 106 Mr ssRNA genome that depends on tobamoviruses for its replication. The complete nucleotide sequence of STMV RNA deduced in the experiments described here was 1059 nucleotides in length. The efficiency of labeling viral RNA with [.gamma.-32P]ATP using T4 polynucleotide kinase was not affected by treatment with tobacco acid pyrophosphatase and/or bacterial alkaline phosphatase, indicating that the majority of the 5'' termini of encapsidated STMV RNAs were not phosphorylated. The 240 3''-terminal nucleotides of STMV RNA and either tobacco mosaic virus (TMV) U1 RNA or TMV U2/U5 RNA had greater than 65% overall sequence similarity, with two nearly identical regions of 40 and 50 bases, respectively. There were no other regions of sequence relatedness to TMV RNA. The 19 5''-terminal nucleotide of STMV RNA had greater than 65% sequence similarity with the 16 5''-terminal nucleotides of brome mosaic virus RNA 3 and 50% sequence similarity with the 12 5''-terminal nucleotides of the Q strain of cucumber mosaic virus RNA 3. The first open reading frame (ORF) beginning at base 53 encoded at 6800 Mr protein that corresponded in size to a major in vitro translation product directed by STMV RNA. A second ORF, beginning at nucleotide 163, had the capacity to code for a protein that corresponded in size (17,500 Mr) to the other major in vitro translation product. The first 12 codons of this ORF corresponded to the sequence of the N-terminal amino acids of the capsid protein. Western-blot analysis of the in vitro translation products revealed that the 17,500 Mr protein had the same electrophoretic mobility as the authentic capsid protein; it was also antigenically related to the capsid protein, but the 6800 Mr protein was not. Time course analysis of in vitro translation demonstrated that the 6800 Mr protein was syntheized at the same time as the capsid protein and did not arise by the proteolytic cleavage of a larger precursor polypeptide. These results suggest that the genome of STMV functioned as a polycistronic messenger RNA. It has not been determined if the 6800 Mr protein is synthesized in vivo. STMV RNA had untranslated regions of 52 and 418 nucleotides at its 5'' and 3'' termini respectively.