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首页> 外文期刊>Journal of Virology >Adenovirus type 2 early polypeptides immunoprecipitated by antisera to five lines of adenovirus-transformed rat cells.
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Adenovirus type 2 early polypeptides immunoprecipitated by antisera to five lines of adenovirus-transformed rat cells.

机译：腺病毒2型早期多肽通过抗血清免疫沉淀到五行的腺病毒转化的大鼠细胞。

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We have identified adenovirus type 2 (Ad2)-induced early polypeptides (EPs) and have attempted to determine which EPs are coded by each of the four early gene blocks. [35S]methionine-labeled EPs were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Cycloheximide pretreatment followed by labeling in hypertonic medium (210 to 250 mM NaCl) facilitated the detection of EPs. Seven major (reproducible bands in autoradiograms) EPs were detected with molecular weights of 74,000 (74K), 21K, 19K, 15K, 13.5K, 11.5K, and 11K. Minor (weaker bands) EPs of 55K, 52K, 42K, 18K, 12K, 8.8K, and 8.3K were also often seen. To identify and map the genes for virus-coded EPs, we prepared antisera against five lines of adenovirus-transformed cells that retain different fractions of the viral genome. The lines were F17, 8617, F4, and T2C4 transformed by Ad2 virions and 5RK (clone I) transformed by transfection with the Ad5 HsuI-G fragment (map position 0 to 8). The early gene blocks retained and expressed (in part) as RNA in these cells were as follows: 5RK(I), block 1 (70% of left 8% of genome); F17, block 1; 8617, blocks 1 and 4; F4 blocks 1, 2, and 4; T2C4, blocks 1, 2, 3, and 4. The following major EPs were immunoprecipitated: 15K by all antisera; 53K and 14.5K by F17, T2C4, 8617, and F4 antisera; 11.5K by T2C4, 8617, and F4 antisera; 44K, 42K, 19K, and 13.5K by T2C4 antisera; 11K by 8617 antisera. Minor EPs of 28K, 18K, and 12K were precipitated by all antisera except 5RK(I). The 53K and 15K EPs were precipitated also from Ad2 early infected monkey cells by the F17 antiserum and by sera from hamsters bearing tumors induced by Ad1-simian virus 40. The relationships between some of the immunoprecipitated EPs were investigated by the partial proteolysis procedure. All 53K EPs are the "same" (i.e., highly related), all 15K EPs are the "same," and all 11.5K EPs are the "same." The 15K EP is highly related to the 14.5 K EP. Although less certain, all 28K EPs appeared related, as did all 18K EPs. The T2C4-specific 44K EP is probably a dimer of the 21K glycopolypeptide. The T2C4-specific 13.5K EP and the 8617-specific 11K EP appear unrelated to any other polypeptides. These immunoprecipitation data provide evidence that early gene block I (map position 1 to 11) may encode major 53K, 15K, and 14.5K polypeptides, and minor 28K, 18K, and 12K polypeptides, and that all or some of the gene for 15K and 14.5K lies within map position 1 to 8. The surprisingly complex pattern of polypeptides coded by early gene block I raises the possibility that some polypeptides may be coded by overlapping "spliced" mRNA's. The possible block locations of the genes for the 21K, 13.5K, and 11.5K polypeptides are discussed.
机译：我们已经鉴定了腺病毒2型（AD2） - 诱导的早期多肽（EPS），并试图确定四种早期基因块中的每一个编码哪些EP。 [35S]通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳分解甲硫氨酸标记的EP。环己酰亚胺预处理，然后在高渗培养基中标记（210至250mM NaCl），促进了EPS的检测。 74,000（74K），21K，19K，15K，13.5K，11.5K和11K的分子量检测到七个主要（放射仪中的可重新发射条带）EPS。还经常看到45k，52k，42k，18k，12k，8.8k和8.3k的45k，52k，42k，18k，12k，8.8k和8.3k的EPS。为了鉴定和映射病毒编码的EPS基因，我们将抗血清对五行的腺病毒转化细胞进行，其保留病毒基因组的不同部分。该线是由Ad2病毒病毒粒子和5RK（克隆I）转化的T 2C 4，通过与AD5 HSUI-G片段转染（Map位置0至8）转化。保留和表达（部分）作为这些细胞中的RNA的早期基因嵌段如下：5RK（I），嵌段1（剩下8％的基因组的70％）; F17，块1; 8617，块1和4; F4块1,2和4; T2C4，块1,2,3和4.以下主要EPS是免疫沉淀的：所有抗血清15K; F17，T2C4,8617和F4抗血清53K和14.5K; 11.5K由T2C4,8617和F4抗血清; 44k，42k，19k和13.5k由T2C4抗血清; 11k到8617抗血清。除了5RK（i）之外，所有抗血清都沉淀了28k，18k和12k的次要EPS。来自AD2早期感染的猴细胞和来自携带Ad1-Simian病毒40诱导的肿瘤的兔血清，53K和15K的EPS沉淀出来。通过部分蛋白水解方法研究了一些免疫沉淀的EPS之间的关系。所有53K eps都是“相同”（即，高度相关），所有15K的EPS都是“相同”，所有11.5K EPS都是“相同”。 15K EP与14.5 K EP有高度相关。虽然少了，但所有28K的EPS都出现了相关的，这是所有18K的EPS。 T2C4特异性44K EP可能是21K甘型肽的二聚体。 T2C4特异性的13.5K EP和8617个特异性11K EP与任何其他多肽无关。这些免疫沉淀数据提供了证据，即早期基因块I（地图位置1至11）可以编码主要的53K，15K和14.5K多肽，并轻微28K，18K和12K多肽，以及15K和15K的全部或一些基因14.5k位于地图位置1至8内。由早期基因嵌段编码的多肽的令人惊讶的复杂模式提出了可以通过重叠的“剪接”mRNA来编码一些多肽的可能性。讨论了21K，13.5K和11.5K多肽基因的可能块位置。

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    《Journal of Virology》 |1979年第1期|共14页
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    W S Wold; M Green;

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  • 收录信息 美国《科学引文索引》(SCI);
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