Mapping of multiple phosphorylation sites within the structural and catalytic domains of the Fujinami avian sarcoma virus transforming protein
Mapping of multiple phosphorylation sites within the structural and catalytic domains of the Fujinami avian sarcoma virus transforming protein
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富士南禽肉瘤病毒转化蛋白结构域和催化域内多个磷酸化位点的定位
作者：
G. Weinmaster;E. Hinze;T. Pawson
福建禽流感肉瘤病毒P140gag-fps基因产物的磷酸化位点已被确定并定位于该转化蛋白的不同区域。从转化细胞中分离到的FSV P140gag-fps至少有三个不同的酪氨酸残基和一个丝氨酸残基磷酸化，此外还有与Pr76gag共享的次要磷酸化位点。用病毒粒子蛋白酶p15或金黄色葡萄球菌V8蛋白酶进行部分蛋白水解已被用于生成P140gag-fps的确定肽片段，从而绘制其磷酸化位点。P140gag-fps的氨基末端gag编码区除了正常的gag磷酸化位点外，还含有一个磷酸酪氨酸残基。两个主要的磷酸酪氨酸残基和主要的磷酸丝氨酸残基位于P140gag-fps编码区的羧基末端。在体外免疫复合物激酶反应中放射性标记的P140gag-fps在体内仅在两个c端酪氨酸残基中的一个磷酸化，在gag编码的酪氨酸和在体内未检测到磷酸化的酪氨酸位点弱磷酸化。因此，P140gag-fps的体外酪氨酸磷酸化与在转化细胞中观察到的不同。对三种福建禽流感肉瘤病毒变异gag-fps蛋白的比较色氨酸肽分析表明，含有磷酸酪氨酸的肽是不变的，这种高度的序列保守性表明这些位点在功能上很重要或位于重要区域。PRCII型禽肉瘤病毒的P105gag-fps转化蛋白缺少福建禽肉瘤病毒P140gag-fps的一个c端磷酸酪氨酸位点。用抗gag血清免疫沉淀的FSV P140gag-fps的部分胰蛋白酶裂解释放免疫复合物中45K和29K的c端片段，保留相关酪氨酸特异性蛋白激酶活性。这一观察结果，以及P140gag-fps的主要磷酸化位点定位于C端fps区域，表明P140gag-fps的激酶结构域位于其C端。P140gag-fps本身的磷酸化是复杂的，这表明它本身可能与转化细胞中的几种蛋白激酶相互作用。
The phosphorylation sites of the P140gag-fps gene product of Fujinami avian sarcoma virus have been identified and localized to different regions of this transforming protein. FSV P140gag-fps isolated from transformed cells is phosphorylated on at least three distinct tyrosine residues and one serine residue, in addition to minor phosphorylation sites shared with Pr76gag. Partial proteolysis with virion protease p15 or with Staphylococcus aureus V8 protease has been used to generate defined peptide fragments of P140gag-fps and thus to map its phosphorylation sites. The amino-terminal gag-encoded region of P140gag-fps contains a phosphotyrosine residue in addition to normal gag phosphorylation sites. The two major phosphotyrosine residues and the major phosphorserine residue are located in the carboxy-terminal portion of the fps-encoded region of P140gag-fps. P140gag-fps radiolabeled in vitro in an immune complex kinase reaction is phosphorylated at only one of the two C-terminal tyrosine residues phosphorylated in vivo and weakly phosphorylated at the gag-encoded tyrosine and at a tyrosine site not detectably phosphorylated in vivo. Thus, the in vitro tyrosine phosphorylation of P140gag-fps is distinct from that seen in the transformed cell. A comparative tryptic phosphopeptide analysis of the gag-fps proteins of three Fujinami avian sarcoma virus variants showed that the phosphotyrosine-containing peptides are invariant, and this high degree of sequence conservation suggests that these sites are functionally important or lie within important regions. The P105gag-fps transforming protein of PRCII avian sarcoma virus lacks one of the C-terminal phosphotyrosine sites found in Fujinami avian sarcoma virus P140gag-fps. Partial trypsin cleavage of FSV P140gag-fps immunoprecipitated with anti-gag serum releases C-terminal fragments of 45K and 29K from the immune complex that retain an associated tyrosine-specific protein kinase activity. This observation, and the localization of the major P140gag-fps phosphorylation sites to the C-terminal fps region, indicate that the kinase domain of P140gag-fps is located at its C terminus. The phosphorylation of P140gag-fps itself is complex, suggesting that it may itself interact with several protein kinases in the transformed cell.
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新型禽肉瘤病毒PRCII的致病性和缺陷。
影响因子：
3.7
作者：
Breitman,ML;Neil,JC;Moscovici,C;Vogt,PK
通讯作者：
Vogt,PK
转化逆转录病毒的基因。
DOI：
10.1101/sqb.1980.044.01.005
发表时间：
1980
期刊：
Cold Spring Harbor symposia on quantitative biology
影响因子：
--
作者：
Duesberg,PH
通讯作者：
Duesberg,PH
禽肉瘤病毒和猫肉瘤病毒的转化序列之间存在同源性。
DOI：
10.1073/pnas.77.11.6536
发表时间：
1980
影响因子：
11.1
作者：
Shibuya,M;Hanafusa,T;Hanafusa,H;Stephenson,JR
通讯作者：
Stephenson,JR
与禽肉瘤病毒 PRCII 的 p105 相关的酪氨酸特异性蛋白激酶活性。
影响因子：
3.7
作者：
Neil,JC;Ghysdael,J;Vogt,PK
通讯作者：
Vogt,PK
三类禽肉瘤病毒编码的蛋白质的结构相似性。
影响因子：
3.7
作者：
Neil,JC;Ghysdael,J;Smart,JE;Vogt,PK
通讯作者：
Vogt,PK