CHARACTERIZATION OF A VARIANT VIRUS SELECTED IN RAT BRAINS AFTER INFECTION BY CORONAVIRUS MOUSE HEPATITIS-VIRUS JHM
CHARACTERIZATION OF A VARIANT VIRUS SELECTED IN RAT BRAINS AFTER INFECTION BY CORONAVIRUS MOUSE HEPATITIS-VIRUS JHM
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DOI：
10.1128/jvi.54.2.429-435.1985
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发表时间：
1985-01-01
影响因子：
5.4
通讯作者：
TERMEULEN, V
中科院分区：
文献类型：
--
作者：
TAGUCHI, F;SIDDELL, SG;TERMEULEN, V
Lewis 大鼠脑内接种鼠冠状病毒小鼠肝炎病毒 (MHV)-JHM 会导致大多数动物患上急性脑炎并在 14 天内死亡。通过Northern印迹分析将感染后5-7天从动物脑中分离的病毒RNA与野生型MHV-JHM（wt病毒）裂解感染Sac（-）或DBT细胞期间产生的RNA进行比较。可重复地，在脑源性材料中，亚基因组 mRNA 2 和 3 但没有其他病毒 RNA 明显更大。当在 Sac(-) 或 DBT 细胞中培养时，从受感染的脑材料中分离出的所有病毒都显示并维持了这种改变的 mRNA 谱。进一步鉴定了从受感染脑材料中分离出的病毒 MHV-JHM 克隆 2（cl-2 病毒）。该分离株在组织培养物中生长并诱导与wt病毒诱导的细胞病变效应相当的细胞病变效应。 cl-2病毒感染的细胞中产生的mRNA 2和3具有大约MW。比感染wt病毒的细胞中产生的多150,000。通过琼脂糖凝胶电泳测定，基因组大小的 RNA (mRNA 1) 或亚基因组 mRNA 4、5、6 和 7 没有可检测到的差异。基因组 RNA 的 T1 抗性寡核苷酸分析显示，与 wt 病毒相比，cl-2 病毒的指纹中多了 1 个寡核苷酸，1 个缺失了寡核苷酸。两种病毒的胞内 mRNA 3 的寡核苷酸指纹是相同的。在衣霉素存在下用[35S]甲硫氨酸脉冲标记显示mRNA 3的初级翻译产物，即E2脱辅基蛋白，是.apprx。 cl-2 病毒感染细胞的 MW 增加了 15,000。这些数据表明，具有较大 mRNA 2 和 3（后者编码改变的 E2 糖蛋白）的病毒被选择在大鼠大脑中进行增殖。考虑了此类变体的产生机制及其选择优势的可能性质。
The intracerebral inoculation of Lewis rats with the murine coronavirus mouse hepatitis virus (MHV)- JHM leads in the majority of animals to acute encephalitis and death within 14 days. Viral RNA isolated from the brains of animals 5-7 days after infection were compared by Northern blot analysis with the RNA produced during the lytic infection of Sac(-) or DBT cells with wild-type MHV-JHM (wt virus). Reproducibly, the subgenomic mRNA 2 and 3 but no other viral RNA were significantly larger in the brain-derived material. All viruses isolated from infected brain material displayed and maintained this altered mRNA profile when cultivated in Sac(-) or DBT cells. A virus isolated from the infected brain material, MHV-JHM clone 2 (cl-2 virus), was further characterized. This isolate grew in tissue culture and induced cytopathic effects comparable to those induced by wt virus. The mRNA 2 and 3 produced in cl-2 virus-infected cells had MW .apprx. 150,000 larger than those produced in cells infected with wt virus. There was no detectable difference in genome-sized RNA (mRNA 1) or subgenomic mRNA 4, 5, 6 and 7 as determined by electrophoresis in agarose gels. T1-resistant oligonucleotide analysis of genomic RNA revealed 1 additional and 1 missing oligonucleotide in the fingerprint of cl-2 virus compared with wt virus. The oligonucleotide fingerprints of intracellular mRNA 3 were identical for both viruses. Pulse-labeling with [35S]methionine in the presence of tunicamycin showed that the primary translation product of mRNA 3, the E2 apoprotein, was .apprx. 15,000 larger in MW in cl-2 virus-infected cells. These data show that viruses with larger mRNA 2 and 3 (the latter encoding an altered E2 glycoprotein) are selected for multiplication in rat brains. Mechanisms for the generation of such variants and the possible nature of their selective advantage are considered.