Overlapping sets of viral RNAs reflect the array of polypeptides in the EcoRI J and N fragments (map positions 81.2 to 85.0) of the Autographa californica nuclear polyhedrosis virus genome
Overlapping sets of viral RNAs reflect the array of polypeptides in the EcoRI J and N fragments (map positions 81.2 to 85.0) of the Autographa californica nuclear polyhedrosis virus genome
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重叠的病毒 RNA 组反映了苜蓿银纹夜蛾核型多角体病毒基因组的 EcoRI J 和 N 片段（图谱位置 81.2 至 85.0）中的多肽阵列
DOI：
10.1128/jvi.61.10.3048-3057.1987
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发表时间：
1987
影响因子：
5.4
通讯作者：
W. Doerfler
中科院分区：
文献类型：
--
作者：
C. Oellig;B. Happ;T. Muller;W. Doerfler
在苜蓿银纹夜蛾核型多角体病毒（Autographa californica nuclear polyhedrosis virus，AcNPV）基因组的几个部分中，已经识别出具有共同3'或5'末端的重叠RNA的嵌套集合。在本报告中，病毒的转录模式和病毒基因产物的安排在81.2至85.0地图单位的区域进行了研究。在AcNPV基因组的这一区段中，鉴定出至少9种大小类别的病毒RNA，其大小范围为1.3 kb至4.6 kb，并显示出共同的3'末端。测定了AcNPV基因组这一部分的酶切图谱和核苷酸序列。计算机分析显示，在右侧转录链上有几个开放阅读框（ORF），在许多潜在的ORF和一些定位RNA的5'末端之前有潜在的TATA和CAAT信号。向左转录的链缺乏主要的ORF。由较大ORF编码的推定多肽的大小范围为11.3至55.6千道尔顿（kDa）。推定的多肽编码的ORF 3，一个33.6 kDa的分子的氨基酸序列，表现出不寻常的，成簇的16倍重复的二肽丝氨酸-丝氨酸的蛋白质，表现出总体优势的碱性氨基酸。与81.2- 85.0-图谱单位区域的内部亚片段同源的杂交选择RNA的体外翻译实验的结果产生约28、34 - 36和48 - 50 kDa的多肽，其大小接近于来自核苷酸序列的主要ORF的长度。通过S1保护分析，在病毒基因组的81.2- 85.0-图谱单位区域中，在3'和5'末端精确确定了RNA的单个大小类别的定位。在8个核苷酸的序列内，所有RNA具有相同的3'末端，其靠近多个聚腺苷酸化信号。精确地绘制了9种不同RNA大小类别的起始位点。当通过S1保护分析确定较小RNA（小于1.8kb）的帽位点时，大量RNA起始位点变得明显。它还表明，在81.2至85.0图单位区域的不同RNA大小的类是可检测的，早在2小时，至少直到36至48小时感染后。在大肠杆菌胞质RNA中，对EcoRI J片段特异的病毒RNA的大小类别在感染后的早期和晚期均可检测到，尽管在早期可检测到较小数量的较大RNA。（摘要截短至400字）
In several parts of the Autographa californica nuclear polyhedrosis virus (AcNPV) genome, nested sets of overlapping RNAs with common 3' or 5' termini have been recognized. In the present report, the pattern of viral transcription and the arrangement of viral gene products in the region of 81.2 to 85.0 map units were investigated. In this segment of the AcNPV genome, at least nine size classes of viral RNA were identified which ranged in size from 1.3 kilobases (kb) to 4.6 kb and exhibited common 3' termini. The detailed restriction map and the nucleotide sequence of this part of the AcNPV genome were determined. Computer analyses revealed several open reading frames (ORFs) on the rightward-transcribed strand with potential TATA and CAAT signals preceding many of the potential ORFs and the 5' termini of some of the mapped RNAs. The leftward-transcribed strand was devoid of major ORFs. The presumptive polypeptides encoded by the larger ORFs ranged in size from 11.3 to 55.6 kilodaltons (kDa). The amino acid sequence of the presumptive polypeptide encoded by ORF3, a 33.6-kDa molecule, exhibited an unusual, clustered 16-fold repeat of the dipeptide arginine-serine in a protein that showed an overall preponderance of basic amino acids. The results of in vitro translation experiments with hybrid-selected RNAs homologous to internal subfragments of the 81.2- to 85.0-map-unit region yielded polypeptides of approximately 28, 34 to 36, and 48 to 50 kDa, which were close in size to the lengths of the major ORFs derived from the nucleotide sequence. The localizations of individual size classes of RNAs in the 81.2- to 85.0-map-unit region of the viral genome were determined precisely at the 3' and 5' termini by S1 protection analyses. Within a sequence of eight nucleotides, all RNAs had the same 3' terminus, which lay close to multiple polyadenylation signals. The initiation sites of the nine different RNA size classes were precisely mapped. As the cap sites of the smaller RNAs (less than 1.8 kb) were determined by S1 protection analyses, a multitude of RNA initiation sites became apparent. It was also shown that the different RNA size classes in the 81.2- to 85.0-map-unit region were detectable as early as 2 h and at least until 36 to 48 h after infection. In unselected cytoplasmic RNA, the size classes of viral RNAs specific for the EcoRI J fragment were detectable early as well as late after infection, although at early times the larger RNAs were detectable in smaller amounts.(ABSTRACT TRUNCATED AT 400 WORDS)