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劳氏肉瘤病毒介导的转化在鸡胚皮肤成纤维细胞和椎体软骨细胞中诱导出相似的糖胺聚糖合成模式。

Transformation by Rous sarcoma virus induces similar patterns of glycosaminoglycan synthesis in chick embryo skin fibroblasts and vertebral chondroblasts.

作者信息

Shanley D J, Cossu G, Boettiger D, Holtzer H, Pacifici M

出版信息

J Biol Chem. 1983 Jan 25;258(2):810-6.

PMID:6296083
Abstract

Chick embryo skin fibroblasts and vertebral chondroblasts were infected with a temperature-sensitive mutant of Rous sarcoma virus, LA24A, and were grown at permissive (36 degrees C) and nonpermissive (41 degrees C) temperatures. During exponential growth, infected and parallel uninfected cultures were labeled with D-[3H]glucosamine, and newly synthesized glycosaminoglycans were identified by anion exchange chromatography and by selective enzymatic and chemical degradations. Control fibroblasts synthesized low levels of hyaluronic acid (HA), and dermatan sulfate (DS), moderate levels of heparan sulfate (HS), and high levels of chondroitin sulfate (CS). In contrast, control chondroblasts synthesized very low levels of HA and DS, no HS, and very high levels of CS. Following transformation and growth at 36 degrees C, both cell types showed a dramatic increase in HA synthesis and a significant decrease in CS synthesis. In addition, transformed chondroblasts initiated the synthesis of HS and increased their synthesis of DS to levels that matched those of transformed fibroblasts. The CS chains synthesized by control chondroblasts were partially undersulfated, while those synthesized by both normal and transformed fibroblasts were fully sulfated. Upon transformation, chondroblasts grown at 36 degrees C initiated the synthesis of fully sulfated CS chains. Most of the above biosynthetic alterations were completely reversed when infected cells were grown at 41 degrees C, indicating that they were dependent on the transforming gene product of LA24A. Clearly, the profound differences that distinguish normal fibroblasts from normal chondroblasts are lost upon transformation, and these two types of terminally differentiated cells converge toward a common, though not identical, biosynthetic program for glycosaminoglycans.

摘要

将鸡胚皮肤成纤维细胞和椎体软骨细胞用劳氏肉瘤病毒的温度敏感突变体LA24A进行感染,并在允许温度(36摄氏度)和非允许温度(41摄氏度)下培养。在指数生长期,对感染的和平行的未感染培养物用D-[3H]葡糖胺进行标记,新合成的糖胺聚糖通过阴离子交换色谱以及选择性酶解和化学降解来鉴定。对照成纤维细胞合成低水平的透明质酸(HA)和硫酸皮肤素(DS)、中等水平的硫酸乙酰肝素(HS)以及高水平的硫酸软骨素(CS)。相比之下,对照软骨细胞合成极低水平的HA和DS,不合成HS,合成非常高水平的CS。在36摄氏度下转化并生长后,两种细胞类型的HA合成均显著增加,CS合成显著减少。此外,转化的软骨细胞开始合成HS,并将其DS合成增加到与转化的成纤维细胞相当的水平。对照软骨细胞合成的CS链部分硫酸化不足,而正常和成纤维细胞合成的CS链均完全硫酸化。在转化后,在36摄氏度下生长的软骨细胞开始合成完全硫酸化的CS链。当感染的细胞在41摄氏度下生长时,上述大多数生物合成改变完全逆转,这表明它们依赖于LA24A的转化基因产物。显然,正常成纤维细胞与正常软骨细胞之间的显著差异在转化后消失,这两种终末分化细胞趋向于一种共同的、虽不完全相同的糖胺聚糖生物合成程序。

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