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12型腺病毒转化区的mRNA种类和蛋白质:体外和体内从1B区2.2kb mRNA的多个编码片段翻译的蛋白质的鉴定

mRNA species and proteins of adenovirus type 12 transforming regions: identification of proteins translated from multiple coding stretches in 2.2 kb region 1B mRNA in vitro and in vivo.

作者信息

Saito I, Shiroki K, Shimojo H

出版信息

Virology. 1983 Jun;127(2):272-89. doi: 10.1016/0042-6822(83)90143-5.

Abstract

Analysis of cytoplasmic RNA in Ad12-infected and -transformed cells showed that more than 20 mRNA species are transcribed from regions 1A and 1B. These mRNA species could be classified into four classes according to the extent to which they were expressed in infected and transformed cells under various conditions. The proteins synthesized from the major species of each of these mRNAs were identified by in vitro translation. In region 1A, the 40K (and 38K) proteins were synthesized from three mRNA species that differed from one another in their 5' ends, and in region 1B three proteins with molecular weights of 19,000 (19K), 50K, and 17K were synthesized from a single species of 2.2 kb mRNA. Only the 19K protein was immunoprecipitated by anti-T serum (G serum) from rats bearing GY1 cell tumors, [GY1 cells are rat cells transformed by the Ad12 HindIII-G fragment (0-6.8 map units).] The 19K protein was also detected by immunoprecipitation in extracts from both transformed and infected cells. The 17K protein was immunoprecipitated by anti-T serum (C serum) derived from rats bearing CY1 cell tumors, but was not by G serum. [CY1 cells are rat cells transformed by the Ad12 EcoRI-C fragment (0-16.5 map units).] The 17K protein was also translated in vitro from the 0.5 kb region 1B mRNA. These results suggest that the 19K and 17K proteins correspond to Ad12 T antigen f and polypeptide IX, respectively, and that these two proteins are translated in vivo from different coding regions in 2.2 kb mRNA in CY1 cells.

摘要

对腺病毒12型(Ad12)感染和转化细胞中的细胞质RNA分析表明,超过20种mRNA种类是从1A和1B区域转录而来的。根据这些mRNA种类在不同条件下在感染和转化细胞中的表达程度,可将它们分为四类。通过体外翻译鉴定了由这些mRNA的主要种类所合成的蛋白质。在1A区域,40K(和38K)蛋白质是由三种在5'端彼此不同的mRNA种类合成的,在1B区域,分子量为19,000(19K)、50K和17K的三种蛋白质是由一种2.2 kb的mRNA合成的。只有19K蛋白质能被来自携带GY1细胞瘤的大鼠的抗T血清(G血清)免疫沉淀,[GY1细胞是被Ad12 HindIII - G片段(0 - 6.8图谱单位)转化的大鼠细胞。]在转化细胞和感染细胞的提取物中,通过免疫沉淀也检测到了19K蛋白质。17K蛋白质能被来自携带CY1细胞瘤的大鼠的抗T血清(C血清)免疫沉淀,但不能被G血清免疫沉淀。[CY1细胞是被Ad12 EcoRI - C片段(0 - 16.5图谱单位)转化的大鼠细胞。]17K蛋白质也能从0.5 kb的1B区域mRNA在体外翻译而来。这些结果表明,19K和17K蛋白质分别对应于Ad12 T抗原f和多肽IX,并且这两种蛋白质在CY1细胞中是从2.2 kb mRNA中的不同编码区域在体内翻译而来的。

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