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在两个持续感染的HeLa细胞系中麻疹病毒特异性多肽的合成

Measles virus-specified polypeptide synthesis in two persistently infected HeLa cell lines.

作者信息

Wechsler S L, Rustigian R, Stallcup K C, Byers K B, Winston S H, Fields B N

出版信息

J Virol. 1979 Sep;31(3):677-84. doi: 10.1128/JVI.31.3.677-684.1979.

Abstract

Measles virus-directed protein synthesis was examined in two HeLa cell lines (K11 and K11A) that are persistently infected with wild-type measles virus. Four viral proteins (H, hemagglutination protein; P, nucleocapsid-associated protein; NP, the major nucleocapsid protein; and M, the matrix protein) were readily detected in both cell lines by immune precipitation of [(35)S]methionine-labeled cell extracts followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, three (H, NP, and M) of the four viral proteins in both K11 and K11A cells differed from the corresponding viral proteins synthesized in HeLa cells acutely infected with the parental wild-type virus. In addition, the M protein from K11A cells migrated significantly more slowly on sodium dodecyl sulfate-polyacrylamide gel electrophoresis than the M protein from K11 cells, and there appeared to be slight differences in the H and NP proteins between these two persistently infected cell lines. The altered viral proteins detected in K11 and K11A cells appeared to be the result of viral mutations rather than changes in the host cell, since virus recovered from these cells directed the synthesis of similar aberrant viral proteins in HeLa cells. Virus recovered from K11 cells and virus recovered from K11A cells were both temperature sensitive and grew more slowly than wild-type virus. HeLa cells infected with virus recovered from K11 cells readily became persistently infected, resembling the original persistently infected K11 cells. Thus, viral mutations are associated with persistent measles virus infections in cell cultures.

摘要

在两个持续感染野生型麻疹病毒的HeLa细胞系(K11和K11A)中检测了麻疹病毒指导的蛋白质合成。通过对[(35)S]甲硫氨酸标记的细胞提取物进行免疫沉淀,然后进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳,在这两个细胞系中均易于检测到四种病毒蛋白(H,血凝素蛋白;P,核衣壳相关蛋白;NP,主要核衣壳蛋白;M,基质蛋白)。当通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析时,K11和K11A细胞中四种病毒蛋白中的三种(H、NP和M)与急性感染亲本野生型病毒的HeLa细胞中合成的相应病毒蛋白不同。此外,K11A细胞的M蛋白在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上的迁移速度明显慢于K11细胞的M蛋白,并且这两个持续感染的细胞系之间的H和NP蛋白似乎存在细微差异。在K11和K11A细胞中检测到的改变的病毒蛋白似乎是病毒突变的结果,而不是宿主细胞的变化,因为从这些细胞中回收的病毒在HeLa细胞中指导合成类似的异常病毒蛋白。从K11细胞中回收的病毒和从K11A细胞中回收的病毒均对温度敏感,并且生长速度比野生型病毒慢。用从K11细胞中回收的病毒感染的HeLa细胞很容易变成持续感染,类似于原始的持续感染的K11细胞。因此,病毒突变与细胞培养中持续的麻疹病毒感染有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c2d0/353496/801e8c8950ab/jvirol00189-0110-a.jpg

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