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首页> 外文期刊>Journal of Virology >Adenovirus early region 1B 58,000-dalton tumor antigen is physically associated with an early region 4 25,000-dalton protein in productively infected cells.
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Adenovirus early region 1B 58,000-dalton tumor antigen is physically associated with an early region 4 25,000-dalton protein in productively infected cells.

机译：腺病毒早期地区1b 58,000-dalton肿瘤抗原与高产细胞中的早期区域4 25,000-dalton蛋白有关。

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摘要

In soluble protein extracts obtained from adenovirus productively infected cells, monoclonal antibodies directed against the early region 1B 58,000-dalton (E1B-58K) protein immunoprecipitated, in addition to this protein, a polypeptide of 25,000 molecular weight. An analysis of tryptic peptides derived from this 25K protein demonstrated that it was unrelated to the E1B-58K protein. The tryptic peptide maps of the 25K protein produced in adenovirus 5 (Ad5)-infected HeLa cells and BHK cells were identical, whereas Ad3-infected HeLa cells produced a different 25K protein. The viral origin of this 25K protein was confirmed by an amino acid sequence determination of five methionine residues in two Ad2 tryptic peptides derived from the 25K protein. The positions of these methionine residues in the 25K protein were compared with the nucleotide sequence of Ad2 and uniquely mapped the gene for this protein to early region 4, subregion 6 of the viral genome. A mutant of Ad5 was obtained (Ad5 dl342) which failed to produce detectable levels of the E1B-58K protein. In HeLa cells infected with this mutant, monoclonal antibodies directed against the E1B-58K protein failed to detect the associated 25K protein. In 293 cells infected with Ad5 dl342, which contain an E1B-58K protein encoded by the integrated adenovirus genome, the mutant produced an E4-25K protein which associated with the E1B-58K protein derived from the integrated genome. Extracts of labeled Ad5 dl342-infected HeLa cells (E1B-58K-) were mixed in vitro with extracts of unlabeled Ad5 wild type-infected HeLa cells or 293 cells (E1B-58K+). When the mixed extracts were incubated with the E1B-58K monoclonal antibody, a labeled E4-25K protein was coimmunoprecipitated. When extracts of Ad5 dl342-infected HeLa cells and uninfected HeLa cells (both E1B-58K-) were mixed, the E1B-58K monoclonal antibody failed to immunoselect the E4-25K protein. These data provide evidence that the E1B-58K antigen is physically associated with an E4-25K protein in productively infected cells. This is the same E1B-58K protein that was previously shown to be associated with the cellular p53 antigen in adenovirus-transformed cells.
机译：在从腺病毒的可溶性蛋白质提取物中，从腺病毒产生的细胞中获得的单克隆抗体，针对早期区域1b 58,000-dalton（E1b-58k）蛋白免疫沉淀，除了该蛋白质，分子量为25,000个多肽。衍生自该25K蛋白质的胰蛋白酶肽的分析表明它与E1B-58K蛋白质不相关。在腺病毒5（Ad5） - 摄入的HeLa细胞和BHK细胞中产生的25K蛋白质的胰蛋白酶肽图是相同的，而AD3感染的HeLa细胞产生了不同的25K蛋白质。通过衍生自25K蛋白的两种AD2胰蛋白酶的五种蛋氨酸残基的氨基酸序列确定该25K蛋白的病毒来源。将这些甲硫氨酸残基在25K蛋白质中的位置与Ad2的核苷酸序列进行比较，并且唯一地映射了该蛋白质的基因，至早期区域4，病毒基因组的亚区域6。获得AD5的突变体（AD5 DL342），其未能产生E1B-58K蛋白的可检测水平。在感染该突变体的Hela细胞中，针对E1B-58K蛋白的单克隆抗体未检测到相关的25K蛋白质。在933个细胞中感染含有由集成腺病毒基因组编码的E1B-58K蛋白质的AD5 DL342，突变体产生了与源自综合基因组的E1B-58K蛋白质相关的E4-25K蛋白。将标记的AD5 DL342感染的HeLa细胞（E1b-58k-）的提取物在体外混合，用未标记的Ad5野生型Hela细胞或293个细胞（E1b-58k +）的提取物。将混合提取物与E1B-58K单克隆抗体一起孵育时，标记的E4-25K蛋白是CoImMunopectated。当混合AD5 DL342感染的HeLa细胞和未感染的HeLa细胞（E1B-58K-）的提取时，E1B-58K单克隆抗体未被免疫e4-25K蛋白质。这些数据提供了表现为E1B-58K抗原在有效感染细胞中与E4-25K蛋白质的物理相关。这是与腺病毒转化细胞中的细胞P53抗原相关联的相同的E1B-58K蛋白质。

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