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劳氏肉瘤病毒src基因表达对鸡胚肌管中收缩蛋白合成的选择性作用。

Selective effect of Rous sarcoma virus src gene expression on contractile protein synthesis in chick embryo myotubes.

作者信息

West C M, Boettiger D

出版信息

Cancer Res. 1983 May;43(5):2042-6.

PMID:6299537
Abstract

Myogenic precursor cells were infected with a temperature-sensitive mutant of Rous sarcoma virus and maintained at the permissive temperature for transformation. Following subculture, a population of cells was produced which failed to differentiate at the permissive temperature but produced a high proportion of myotubes in sister cultures shifted to the nonpermissive temperature. The myotube-containing cultures were further enriched for this cell type by the addition of 1-beta-D-arabinofuranosylcytosine to kill replicating mononucleated cells. This myotube population was suitable for testing the effect of viral-transforming gene expression in a postmitotic, terminally differentiated cell which expresses relatively low levels of the cellular homologue of the viral-transforming gene and is resistant to infection by the virus. The consequence of shifting these Rous sarcoma virus-infected myotube cultures to the permissive temperature was assessed in terms of protein synthesis. The total rate of incorporation of exogenous radioactive leucine, supplied at a concentration which saturated the intracellular pool, was similar between cultures held at the two temperatures, suggesting that the absolute rate of total protein synthesis was not affected by expression of viral-transforming gene. In contrast, the rate of synthesis of eight proteins the expression of which is specific for myotubes was suppressed reversibly. The rate of synthesis of five other proteins which are not selectively concentrated in myotubes was either unaffected or stimulated. Thus. the expression of viral-transforming gene in myotubes leads to differential effects on developmentally regulated proteins without inducing several properties of the transformed state classically observed for fibroblastic cells.

摘要

将成肌前体细胞用劳氏肉瘤病毒的温度敏感突变体进行感染,并在允许转化的温度下进行培养。继代培养后,产生了一群细胞,这些细胞在允许温度下不能分化,但在转移至非允许温度的姐妹培养物中能产生高比例的肌管。通过添加1-β-D-阿拉伯呋喃糖基胞嘧啶以杀死正在复制的单核细胞,进一步富集了含有肌管的培养物中的这种细胞类型。这群肌管适合用于测试病毒转化基因表达在有丝分裂后、终末分化细胞中的作用,该细胞表达相对低水平的病毒转化基因的细胞同源物,并且对病毒感染具有抗性。从蛋白质合成方面评估了将这些感染劳氏肉瘤病毒的肌管培养物转移至允许温度的后果。在两种温度下培养的细胞中,以饱和细胞内池的浓度供应的外源放射性亮氨酸的总掺入率相似,这表明总蛋白质合成的绝对速率不受病毒转化基因表达的影响。相反,八种其表达对肌管具有特异性的蛋白质的合成速率被可逆地抑制。另外五种没有选择性地集中在肌管中的蛋白质的合成速率要么未受影响,要么受到刺激。因此,病毒转化基因在肌管中的表达对发育调控蛋白产生了不同的影响,而没有诱导成纤维细胞中经典观察到的转化状态的几种特性。

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