Hammond R W, Crosslin J M
Molecular Plant Pathology Laboratory, USDA-Agricultural Research Service, Beltsville, Maryland 20705, USA.
Virology. 1995 Apr 1;208(1):349-53. doi: 10.1006/viro.1995.1159.
The complete nucleotide sequence of RNA 3 of the PE-5 peach isolate of Prunus necrotic ringspot ilarvirus (PNRSV) was obtained from cloned cDNA. The RNA sequence is 1941 nucleotides and contains two open reading frames (ORFs). ORF 1 consisted of 284 amino acids with a calculated molecular weight of 31,729 Da and ORF 2 contained 224 amino acids with a calculated molecular weight of 25,018 Da. ORF 2 corresponds to the coat protein gene. Expression of ORF 2 engineered into a pTrcHis vector in Escherichia coli results in a fusion polypeptide of approximately 28 kDa which cross-reacts with PNRSV polyclonal antiserum. Analysis of the coat protein amino acid sequence reveals a putative "zinc-finger" domain at the amino-terminal portion of the protein. Two tetranucleotide AUGC motifs occur in the 3'-UTR of the RNA and may function in coat protein binding and genome activation. ORF 1 homologies to other ilarviruses and alfalfa mosaic virus are confined to limited regions of conserved amino acids. The translated amino acid sequence of the coat protein gene shows 92% similarity to one isolate of apple mosaic virus, a closely related member of the ilarvirus group of plant viruses, but only 66% similarity to the amino acid sequence of the coat protein gene of a second isolate. These relationships are also reflected at the nucleotide sequence level. These results in one instance confirm the close similarities observed at the biophysical and serological levels between these two viruses, but on the other hand call into question the nomenclature used to describe these viruses.
从克隆的cDNA中获得了李坏死环斑病毒(PNRSV)PE - 5桃分离株RNA 3的完整核苷酸序列。该RNA序列为1941个核苷酸,包含两个开放阅读框(ORF)。ORF 1由284个氨基酸组成,计算分子量为31729道尔顿,ORF 2包含224个氨基酸,计算分子量为25018道尔顿。ORF 2对应于外壳蛋白基因。将工程化到pTrcHis载体中的ORF 2在大肠杆菌中表达,产生一种约28 kDa的融合多肽,该多肽与PNRSV多克隆抗血清发生交叉反应。对外壳蛋白氨基酸序列的分析揭示了该蛋白氨基末端部分存在一个推定的“锌指”结构域。RNA的3'-UTR中出现两个四核苷酸AUGC基序,可能在外壳蛋白结合和基因组激活中起作用。ORF 1与其他等轴不稳环斑病毒和苜蓿花叶病毒的同源性仅限于保守氨基酸的有限区域。外壳蛋白基因的翻译氨基酸序列与苹果花叶病毒的一个分离株显示出92%的相似性,苹果花叶病毒是植物病毒等轴不稳环斑病毒组的一个密切相关成员,但与第二个分离株的外壳蛋白基因氨基酸序列仅显示出66%的相似性。这些关系在核苷酸序列水平上也有所体现。这些结果一方面证实了这两种病毒在生物物理和血清学水平上观察到的密切相似性,但另一方面也对用于描述这些病毒的命名法提出了质疑。