Clinton G M, Finley-Whelan J
J Cell Biol. 1984 Sep;99(3):788-95. doi: 10.1083/jcb.99.3.788.
Tyrosyl kinase activity in vesicular stomatitis virus (VSV) acquired from host cells that differ in morphology was investigated. VSV grown in baby hamster kidney (BHK) cells with rounded morphology and a high efficiency of colony formation in soft agar (Rous sarcoma virus [RSV]-transformed and suspension BHK cells) was compared with VSV grown in BHK cells with a flattened morphology and lower efficiency of colony formation in soft agar (RSV-infected revertant and control BHK cells). Tyrosyl kinase activity measured with the substrates angiotensin II peptide or casein was found at 7-10-fold higher levels in virus released from the anchorage-independent BHK cells. Most of the VSV-associated tyrosyl kinases acquired from the RSV-transformed BHK cells reacted with antiserum to pp60src, whereas the activity acquired from the suspension BHK cells was unaffected by anti-src serum. The overall levels of tyrosyl kinase in subcellular fractions of the host BHK cells were also measured. Like the VSV released from them, the RSV-transformed cell extracts contained high levels. The suspension cells, however, contained the same low levels of tyrosyl kinase as was found in the control BHK cell extracts. Therefore, tyrosyl kinase was concentrated and acquired by VSV from the anchorage-independent suspension BHK cells. VSV-associated protein kinases acquired from other cell types followed a similar pattern. Tyrosyl kinase levels were high in VSV released from suspension cultures (Chinese hamster ovary and HeLa) and from virally transformed cells (Kirsten murine sarcoma virus-transformed rat kidney cells) and low in VSV released from an anchorage-dependent primary cell culture (chick embryo fibroblasts).
对从形态不同的宿主细胞中获得的水疱性口炎病毒(VSV)的酪氨酸激酶活性进行了研究。将在具有圆形形态且在软琼脂中形成集落效率高的幼仓鼠肾(BHK)细胞(劳斯肉瘤病毒[RSV]转化的和悬浮的BHK细胞)中生长的VSV与在具有扁平形态且在软琼脂中形成集落效率较低的BHK细胞(RSV感染的回复细胞和对照BHK细胞)中生长的VSV进行比较。用血管紧张素II肽或酪蛋白作为底物测定的酪氨酸激酶活性,在从非贴壁依赖性BHK细胞释放的病毒中高出7至10倍。从RSV转化的BHK细胞获得的大多数VSV相关酪氨酸激酶与抗pp60src抗血清反应,而从悬浮BHK细胞获得的活性不受抗src血清的影响。还测量了宿主BHK细胞亚细胞组分中酪氨酸激酶的总体水平。与从它们释放的VSV一样,RSV转化的细胞提取物含有高水平的酪氨酸激酶。然而,悬浮细胞中酪氨酸激酶的水平与对照BHK细胞提取物中的水平相同,均较低。因此,酪氨酸激酶被VSV从非贴壁依赖性悬浮BHK细胞中浓缩并获得。从其他细胞类型获得的VSV相关蛋白激酶遵循类似的模式。从悬浮培养物(中国仓鼠卵巢细胞和HeLa细胞)以及病毒转化细胞(柯斯顿鼠肉瘤病毒转化的大鼠肾细胞)释放的VSV中酪氨酸激酶水平较高,而从贴壁依赖性原代细胞培养物(鸡胚成纤维细胞)释放的VSV中酪氨酸激酶水平较低。