Nucleotide and amino acid sequence analysis of the rotavirus nonstructural RNA-binding protein NS35.
Nucleotide and amino acid sequence analysis of the rotavirus nonstructural RNA-binding protein NS35.
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轮状病毒非结构 RNA 结合蛋白 NS35 的核苷酸和氨基酸序列分析。
DOI：
10.1006/viro.1993.1059
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发表时间：
1993
期刊：
影响因子：
3.7
通讯作者：
Kattoura,M
中科院分区：
文献类型：
--
作者：
Patton,JT;Salter-Cid,L;Kalbach,A;Mansell,EA;Kattoura,M
NS 35是轮状病毒SA 11株基因8编码的碱性蛋白，具有RNA结合活性，是轮状病毒基因组复制所必需的。为了确定NS 35基因及其蛋白产物中的保守区域，我们确定了哺乳动物和禽类轮状病毒Wa、DS 1、SA 11（Patton和Ramig株）、NCDV和Ty-1的NS 35基因的核苷酸序列，并将它们及其推导的氨基酸序列与SA 11（Both株）、OSU和UK的报道进行了比较。结果表明，哺乳动物轮状病毒NS 35基因全长1058-1059个碱基，编码317个氨基酸，序列保守性高达83%。火鸡轮状病毒Ty-1的NS 35基因与哺乳动物轮状病毒的NS 35基因在预测的蛋白质（315个氨基酸）和基因（1042个碱基）的大小方面不同。Ty-1的NS 35与哺乳动物病毒的NS 35具有相对低的氨基酸同源性（52-57%）。NS 35基因的系统发育分析表明，鸟类（Ty-1）和哺乳动物轮状病毒是远亲。NS 35的预测序列的比较表明，所有具有保守的基础结构域的37个氨基酸残基的205-241，可能作为RNA结合结构域。电泳检测表明，NS 35含有一个二硫键，可能位于蛋白质的氨基末端的一半。NS 35基因在核苷酸水平上的比较揭示了两个广泛保守的区域，（i）75-bp（B）序列，包括35-bp的5′-非编码区和NS 35开放阅读框的前30个碱基，和（ii）基因3′-非编码区的28-B序列。NS 35 mRNA的二级结构预测表明，75个碱基的序列可以折叠产生茎双环结构。这种结构可以作为包装信号，用于将NS 35 mRNA分类到复制酶颗粒中。
NS35, a basic protein encoded by gene 8 of SA11 rotavirus, possesses RNA-binding activity and is essential for genome replication. To identify conserved regions in the NS35 gene and its protein product, we determined the nucleotide sequences of the NS35 gene for the mammalian and avian rotaviruses Wa, DS1, SA11 (Patton and Ramig strains), NCDV, and Ty-1 and compared them and their deduced amino acid sequences to those reported for SA11 (Both strain), OSU, and UK. The results indicated that the NS35 genes of the mammalian rotaviruses are 1058-1059 bases in length and encode proteins of 317 amino acids that exhibit high levels of sequence conservation (⩾83%). The NS35 gene of the turkey rotavirus Ty-1 differed from those of the mammalian rotaviruses with respect to size of the predicted protein (315 amino acids) and of the gene (1042 bases). NS35 of Ty-1 exhibited a relatively low degree of amino acid homology (52-57%) with NS35 of the mammalian viruses. Phylogenetic analysis of the NS35 gene indicated that avian (Ty-1) and mammalian rotaviruses are distantly related. Comparison of the predicted sequences of NS35 showed that all possessed a conserved basic domain of 37 amino acids at residues 205-241 that may serve as the RNA-binding domain. Electrophoretic examination showed that NS35 contains a disulfide bond probably located in the amino-terminal half of the protein. Comparison of NS35 genes at the nucleotide level revealed two regions of extensive conservation, (i) a 75-base (b) sequence that includes the 35-base 5′-noncoding region and the first 30 bases of the open reading frame for NS35, and (ii) a 28-b sequence in the 3′-noncoding region of the gene. Secondary structure predictions for the NS35 mRNA suggest that the 75-base sequence can fold to produce a stem double-loop structure. Such a structure may serve as a packaging signal for the assortment of NS35 mRNA into replicase particles.