Cross F R, Hanafusa H
Cell. 1983 Sep;34(2):597-607. doi: 10.1016/0092-8674(83)90392-6.
We have constructed mutants of Rous sarcoma virus expressing p60srcs that are underphosphorylated on serine or tyrosine, by linker insertion or insertion/deletion into cloned Rous sarcoma virus DNA, and recovery of mutant virus by transfection of chicken embryo fibroblasts. Cells infected with mutants whose p60srcs lack the major site of either serine or tyrosine phosphorylation were morphologically transformed and formed colonies in soft agar. The tyrosine kinase activities of the mutant p60srcs measured in vivo and in vitro were close to the wild type activity. Peptide mapping showed that phosphorylation on tyrosine and serine of p60src is independent: the major phosphorylated tyrosine and the major phosphorylated serine can each be phosphorylated in the absence of phosphorylation of the other.
我们通过在克隆的劳氏肉瘤病毒DNA中插入接头或进行插入/缺失操作，构建了表达在丝氨酸或酪氨酸上磷酸化不足的p60srcs的劳氏肉瘤病毒突变体，并通过转染鸡胚成纤维细胞来回收突变病毒。感染了p60srcs缺乏丝氨酸或酪氨酸主要磷酸化位点的突变体的细胞发生了形态转化，并在软琼脂中形成了集落。在体内和体外测量的突变体p60srcs的酪氨酸激酶活性接近野生型活性。肽图谱分析表明，p60src在酪氨酸和丝氨酸上的磷酸化是独立的：主要磷酸化的酪氨酸和主要磷酸化的丝氨酸在彼此未磷酸化的情况下均可被磷酸化。
