COMPLETE NUCLEOTIDE-SEQUENCE OF A CELL CULTURE-ADAPTED VARIANT OF HEPATITIS-A VIRUS - COMPARISON WITH WILD-TYPE VIRUS WITH RESTRICTED CAPACITY FOR INVITRO REPLICATION
COMPLETE NUCLEOTIDE-SEQUENCE OF A CELL CULTURE-ADAPTED VARIANT OF HEPATITIS-A VIRUS - COMPARISON WITH WILD-TYPE VIRUS WITH RESTRICTED CAPACITY FOR INVITRO REPLICATION
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DOI：
10.1016/0042-6822(88)90270-x
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发表时间：
1988-04-01
期刊：
影响因子：
3.7
通讯作者：
LEMON, SM
中科院分区：
文献类型：
--
作者：
JANSEN, RW;NEWBOLD, JE;LEMON, SM
为了确定与甲型肝炎病毒（HAV）适应细胞培养生长相关的分子变化，对HAV HM 175株细胞培养适应变异株（p16 HM 175，第16代体外传代水平）的基因组进行了分子克隆，并测定了该病毒的完整核苷酸序列。与野生型病毒相比，p16 HM 175在猴肾（BS-C-1）细胞中有效复制（每一感染单位约58个含RNA的颗粒，与之相比，每一感染单位约2.4 × 10 - 6个含RNA的颗粒）。105）。p16 HM 175的核苷酸序列揭示了野生型基因组中总共19个突变，包括5个在5“非翻译区的突变，1个在3”非翻译区的突变，和13个预测HAV蛋白质氨基酸序列的8个变化的突变。在衣壳蛋白（VP 2）中仅发生一个氨基酸取代，而其他氨基酸取代涉及蛋白2A、2B、2C、VPg和3Dpol。当将p16病毒的序列与先前报道的独立分离的、细胞培养适应的HM 175病毒变体的序列进行比较时（J. I. Cohen等人，（1987），Proc. Natl. Acad. Sci. USA 84，2497-2501），在RNA的非翻译区中存在三个相同的突变，并且在蛋白质VP 2、2B和3Dpol中存在涉及相同氨基酸的四个突变。这些突变在基因组内的分布表明，RNA复制的变化可能是病毒适应体外生长的首要因素。因此，这些数据有助于理解HAV适应细胞培养的分子基础，并且由于减毒经常伴随着病毒适应细胞培养中的生长，因此可能有益于计划减毒疫苗的开发。
To determine the molecular changes associated with adaptation of hepatitis A virus (HAV) to growth in cell culture, the genome of a cell culture-adapted variant of HM175 strain HAV (p16 HM175, 16th in vitro passage level) was molecularly cloned and the complete nucleotide sequence of the virus was determined. Compared with wild-type virus, p16 HM175 replicates efficiently in monkey kidney (BS-C-1) cells (approximately 58 RNA-containing particles per one infectious unit, compared with 2.4 .times. 105 for wild-type HM175). The nucleotide sequence of p16 HM175 revealed a total of 19 mutations from the wild-type genome, including 5 mutations in the 5'' nontranslated region, 1 mutation in the 3'' nontranslated region, and 13 mutations predicting 8 changes in the amino acid sequences of HAV proteins. Only one amino acid substitution occurred among the capsid proteins (VP2), while others involved proteins 2A, 2B, 2C, VPg, and 3Dpol. When the sequence of p16 virus was compared with that reported previously for an independently isolated, cell culture-adapted variant of HM175 virus (J. I. Cohen et al., (1987), Proc. Natl. Acad. Sci. USA 84, 2497-2501), there were three identical mutations in nontranslated regions of the RNA, and four mutations involving identical amino acids in proteins VP2, 2B, and 3Dpol. The distribution of these mutations within the genome suggests that changes in RNA replication may be of primary importance in adaptation of the virus to growth in vitro. These data are thus helpful in understanding the molecular basis of adaptation of HAV to cell culture and, since attenuation frequently accompanies adaptation of virus to growth in cell culture, may be of benefit in planning for attenuated vaccine development.