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在莫洛尼氏鼠肉瘤病毒转化的细胞中检测一种转化基因产物。

Detection of a transforming gene product in cells transformed by Moloney murine sarcoma virus.

作者信息

Papkoff J, Verma I M, Hunter T

出版信息

Cell. 1982 Jun;29(2):417-26. doi: 10.1016/0092-8674(82)90158-1.

Abstract

We identified, in cells transformed by Moloney murine sarcoma virus (M-MuSV clone 124), a protein encoded by the M-MuSV transforming gene, v-mos. An antiserum against a synthetic peptide corresponding to the C terminus of a protein predicted from the v-mos nucleotide sequence specifically recognizes a protein doublet of approximately 37,000 daltons from 35S-methionine-labeled M-MuSV 124-transformed producer cells. By peptide mapping, this protein is almost identical to the 37 kd in vitro translation product from the M-MuSV v-mos gene. Immunoprecipitates from 32P-labeled cells contain a single v-mos-specific phosphoprotein, which has at least six sites of phosphorylation containing phosphoserine. Pulse-chase experiments show that the lower band in the 35S-methionine-labeled doublet is the primary translation product, which is modified, probably by phosphorylation, to yield the upper band. A similar mos protein is immunoprecipitated from HT1-MuSV-transformed cells, but not from uninfected NIH/3T3 cells. These mos proteins are present at very low levels in transformed cell lines. Cells acutely infected with M-MuSV 124, however, transiently contain much higher levels of the mos protein. These high levels coincide with extensive cell mortality.

摘要

我们在莫洛尼鼠肉瘤病毒(M-MuSV克隆124)转化的细胞中鉴定出一种由M-MuSV转化基因v-mos编码的蛋白质。针对根据v-mos核苷酸序列预测的蛋白质C末端合成肽的抗血清,能特异性识别来自35S-甲硫氨酸标记的M-MuSV 124转化的产生细胞中约37,000道尔顿的蛋白质双峰。通过肽图谱分析,该蛋白质与M-MuSV v-mos基因的37 kd体外翻译产物几乎相同。来自32P标记细胞的免疫沉淀产物包含一种单一的v-mos特异性磷蛋白,其至少有六个含磷酸丝氨酸的磷酸化位点。脉冲追踪实验表明,35S-甲硫氨酸标记双峰中的较低条带是主要翻译产物,可能通过磷酸化修饰产生较高条带。从HT1-MuSV转化的细胞中可免疫沉淀出类似的mos蛋白,但未感染的NIH/3T3细胞中则没有。这些mos蛋白在转化细胞系中的含量非常低。然而,急性感染M-MuSV 124的细胞会短暂含有更高水平的mos蛋白。这些高水平与广泛的细胞死亡同时出现。

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