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首页> 外文期刊>Journal of Virology >Association of pp60src and src protein kinase activity with the plasma membrane of nonpermissive and permissive avian sarcoma virus-infected cells.
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Association of pp60src and src protein kinase activity with the plasma membrane of nonpermissive and permissive avian sarcoma virus-infected cells.

机译：PP60SRC和SRC蛋白激酶活性与非智能和允许禽Sarcoma病毒感染细胞的血浆膜的关联。

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摘要

The intracellular localization of pp60src and src protein kinase activity in avian sarcoma virus (ASV)-infected chicken embryo fibroblasts and transformed and morphologically reverted field vole cells was examined by subcellular fractionation procedures. Fractionation by differential centrifugation of Dounce-homogenized cellular extracts prepared from vole cells showed that 83 to 91% of pp60src sedimented with particulate subcellular components from both transformed and revertant vole cells. A slightly lesser amount (60 to 70%) of pp60src was found associated with the particulate fraction from ASV-infected chicken embryo fibroblasts. The distribution of src protein kinase activity in the cytosol and particulate cell fractions was identical to that of pp60src, indicating no detectable differences in the activity of cytosol- and particulate-associated pp60src. When subcellular components of the cell were fractionated by discontinuous sucrose gradient centrifugation, similar amounts of both pp60src and src protein kinase activity cosedimented with the plasma membrane fractions from both transformed and revertant vole cells, as well as from ASV-infected chicken embryo fibroblasts. src protein kinase activity associated with plasma membrane fractions prepared from vole cells and ASV-infected chicken embryo fibroblasts was resistant to extraction with high salt concentrations, but partial elution was achieved with nonionic detergent. Thus, in both transformed and morphologically reverted vole cells, pp60src is intimately associated with the plasma membrane. Since transforming virus can be rescued from revertant vole cells by fusion to chicken embryo fibroblasts, revertant vole cell pp60src is capable of inducing morphological transformation. Thus, although the data presented herein suggest that transformation requires the association of pp60src with the plasma membrane, the binding of pp60src to the plasma membrane per se is insufficient to induce morphological transformation and requires the additional interaction with a specific target membrane protein which appears to be defective in revertant vole cells.
机译：通过亚细胞分级方法检查PP60SRC和SRC蛋白激酶活性的PP60SRC和SRC蛋白激酶活性的细胞内定位和转化的和形态恢复的野孔细胞。通过沟细胞制备的常规离心的分馏来分馏，显示出83至91％的PP60SRC与来自转化和还原的孔细胞的颗粒状亚细胞组分沉淀。发现与来自ASV感染的鸡胚成纤维细胞的颗粒级分相关的PP60SRC略少的量（60％至70％）。胞浆溶胶和颗粒细胞级分中的SRC蛋白激酶活性的分布与PP60SRC的SRC蛋白激酶活性相同，表明在胞质和颗粒相关的PP60SRC的活性中没有可检测的差异。当细胞的亚细胞组分通过不连续蔗糖梯度离心分馏时，类似量的PP60SRC和SRC蛋白激酶活性与来自转化的和倒剂孔细胞的血浆膜级分，以及来自受影响的鸡胚胚胎成纤维细胞。与由孔细胞和ASV感染的鸡胚成纤维细胞制备的血浆膜级分相关的SRC蛋白激酶活性与高盐浓度的萃取抗性，但通过非离子洗涤剂实现了部分洗脱。因此，在转化和形态学上恢复的血对细胞中，PP60SRC与质膜密切相关。由于转化病毒可以通过融合到鸡胚成纤维细胞来拯救转化剂孔细胞，因此还原剂孔电池PP60SRC能够诱导形态转化。因此，尽管本文所呈列的数据表明，转化需要PP60SRC与血浆膜的关联，但PP60SRC对质膜本身的结合不足以诱导形态转化，并且需要与特定靶膜蛋白的额外相互作用呈现出来在回复孔细胞中有缺陷。

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