Expression of early viral gene products in adenovirus type 12-infected and -transformed cells.
Expression of early viral gene products in adenovirus type 12-infected and -transformed cells.
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早期病毒基因产物在12型腺病毒感染和转化细胞中的表达。
DOI：
10.1099/0022-1317-60-1-99
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
B. Siegmann
中科院分区：
文献类型：
--
作者：
H. Esche;B. Siegmann
我们分析了早期病毒基因产物表达的腺病毒12型（Ad 12）感染的细胞，以及在两个Ad 12转化的仓鼠细胞系，和Ad 12诱导的大鼠肿瘤细胞系的无细胞翻译的病毒特异性RNA，这是通过杂交选择克隆的限制性内切酶片段的病毒DNA。通过一维和二维凝胶电泳分析体外合成的蛋白质。发现E1 A早期区编码的RNA指导至少8种多肽的合成，其表观分子量为10000。重量38K、36K、30K、28K、26K、25K、24K和22K。所有这些蛋白质都相互关联。E1 B特异性RNA指导三种蛋白质的合成：59 K，19 K和17 K。早期区域E2 a编码61 K多肽，其可能代表Ad 12的单链DNA结合蛋白。与区域E3互补的RNA指导16 K蛋白的合成，并且从区域E4转录的RNA指导具有mol.重量20 K、18 K和11.5K。我们已经将一个67 K多肽定位到11至28个图位单位（E2 b）内的区域。由两个Ad 12转化的仓鼠细胞系（T637，HA 12/7）和一个Ad 12诱导的大鼠肿瘤细胞系（RBT 12/3）制备的病毒特异性RNA指导的蛋白质分析表明，早期区域E1和E4在所有三个Ad 12转化的细胞系中表达。从早期区域E2和E3转录的RNA已在T637和RBT 12/3系中检测到。从Ad 12转化的细胞系制备的病毒RNA指导具有mol.重量与来自感染细胞的早期病毒蛋白非常相似。然而，在上述所有三种Ad 12转化的细胞系中，我们发现了指导早期区域E1（34 K）和E4（25 K，24 K）的额外多肽合成的RNA，这些多肽在感染的细胞中未检测到。11和28个图谱单位之间的DNA序列（编码67 K蛋白）在Ad 12转化的细胞中不表达。
We have analysed early viral gene products expressed in adenovirus type 12 (Ad12)-infected cells as well as in two Ad12-transformed hamster cell lines, and Ad12-induced rat tumour cell lines by cell-free translation of virus-specific RNA which was selected by hybridization to cloned restriction endonuclease fragments of virus DNA. Proteins synthesized in vitro were analysed by one- and two-dimensional gel electrophoresis. It was found that RNA encoded by early region E1A directs the synthesis of at least eight polypeptides with apparent mol. wt. 38K, 36K, 30K, 28K, 26K, 25K, 24K and 22K. All these proteins are related to each other. E1B-specific RNA directs the synthesis of three proteins: 59K, 19K and 17K. Early region E2a codes for a 61K polypeptide which probably represents the single-strand DNA-binding protein of Ad12. RNA complementary to region E3 directs the synthesis of a 16K protein, and RNA transcribed from region E4 the synthesis of polypeptides with mol. wt. 20K, 18K and 11.5K. We have mapped a 67K polypeptide into the region within 11 to 28 map units (E2b). The analysis of proteins directed by virus-specific RNAs prepared from two Ad12-transformed hamster cell lines (T637, HA12/7) and one Ad12-induced rat tumour line (RBT12/3) showed that early regions E1 and E4 are expressed in all three Ad12-transformed cell lines. RNA transcribed from early regions E2 and E3 have been detected in lines T637 and RBT12/3. The virus RNA prepared from the Ad12-transformed cell lines directed synthesis of polypeptides with mol. wt. very similar to those of early virus proteins from infected cells. However, in all three Ad12-transformed cell lines mentioned above we have found RNAs which directed the synthesis of additional polypeptides of early regions E1 (34K) and E4 (25K, 24K) not detected in infected cells. The DNA sequence between 11 and 28 map units (coding for the 67K protein) is not expressed in the Ad12-transformed cells.