Sugawara K, Gilead Z, Wold W S, Green M
J Virol. 1977 May;22(2):527-39. doi: 10.1128/JVI.22.2.527-539.1977.
High-titer monospecific antiserum against highly purified adenovirus 2 (Ad2) single-stranded DNA binding protein (DBP) was used to study, by indirect immunofluorescence (IF), the synthesis of DBP in Ad2-infected human cells and adenovirus-transformed rat, hamster, and human cell lines. In infected cells the synthesis of DBP was first detected in the cytoplasm at 2 to 4 h postinfection and reached a maximum intensity at 6 h postinfection. At this time DBP began to accumulate in the nucleus, where it reached maximum intensity at about 14 h postinfection. The cytoplasmic IF was diffuse, whereas nuclear IF appeared as dots that coalesced into large globules as infection progressed. In cells treated with 1-beta-d-arabinofuranosylcytosine to inhibit viral DNA synthesis, strong nuclear IF was observed in the form of dots, but the large fluorescent globules were not observed. The Ad2 (oncogenic group C) anti-DBP serum reacted very strongly by IF with Ad5 (group C)-infected, to a lesser extent with Ad7 and Ad11 (group B)-infected, and weakly with Ad12 and Ad18 (group A)-infected KB cells (treated with 1-beta-d-arabinofuranosylcytosine). These results may indicate that Ad2 DBP is closely related immunologically to DBPs induced early after infection by adenovirus serotypes in oncogenic group C, moderately related to DBPs of serotypes in oncogenic group B, and perhaps distantly related to DBPs of serotypes in oncogenic group A. The following adenovirus-transformed cell lines were examined for DBP synthesis by IF with the Ad2 anti-DBP serum: six rat cell lines (T2C4, F17, 8662, 8638, 8617, and F161) transformed by Ad2 virus, three hamster cell lines transformed by Ad2 virus (Ad2HT1) and Ad2-simian virus 40 hybrid virus (ND1HK1 and ND4HK4), and one rat (5RK) and one human (293-31) cell line transformed by transfection with Ad5 DNA. T2C4 and 8662 appeared weakly positive, whereas Ad2HT1 and ND4HK1 were strongly positive. The other transformed cell lines did not produce DBP detectable by IF. Thus, some but not all transformed cell lines produce DBP, which indicates that DBP is not required for maintenance of cell transformation and that transformed cells can express "nontransforming" viral genes as protein.
使用针对高度纯化的腺病毒2（Ad2）单链DNA结合蛋白（DBP）的高效价单特异性抗血清，通过间接免疫荧光法（IF）研究Ad2感染的人细胞以及腺病毒转化的大鼠、仓鼠和人细胞系中DBP的合成。在感染的细胞中，感染后2至4小时在细胞质中首次检测到DBP的合成，并在感染后6小时达到最大强度。此时DBP开始在细胞核中积累，并在感染后约14小时达到最大强度。细胞质中的免疫荧光呈弥漫性，而细胞核中的免疫荧光则表现为点状，随着感染的进展，这些点状会合并成大的球状体。在用1-β-D-阿拉伯呋喃糖基胞嘧啶处理以抑制病毒DNA合成的细胞中，观察到强烈的细胞核免疫荧光呈点状，但未观察到大型荧光球状体。Ad2（致癌性C组）抗DBP血清通过免疫荧光法与Ad5（C组）感染的细胞反应非常强烈，与Ad7和Ad11（B组）感染的细胞反应较弱，与Ad12和Ad18（A组）感染的KB细胞（用1-β-D-阿拉伯呋喃糖基胞嘧啶处理）反应微弱。这些结果可能表明，Ad2 DBP在免疫上与致癌性C组腺病毒血清型感染后早期诱导的DBP密切相关，与致癌性B组血清型的DBP中度相关，可能与致癌性A组血清型的DBP关系较远。使用Ad2抗DBP血清通过免疫荧光法检测了以下腺病毒转化细胞系中DBP的合成：六个由Ad2病毒转化的大鼠细胞系（T2C4、F17、8662、8638、8617和F161），三个由Ad2病毒（Ad2HT1）和Ad2-猴病毒40杂交病毒（ND1HK1和ND4HK4）转化的仓鼠细胞系，以及一个通过用Ad5 DNA转染转化的大鼠（5RK）和一个人（293-31）细胞系。T2C4和8,662呈弱阳性，而Ad2HT1和ND/HK1呈强阳性。其他转化细胞系未产生可通过免疫荧光检测到的DBP。因此，一些但不是所有的转化细胞系产生DBP，这表明维持细胞转化不需要DBP，并且转化细胞可以将“非转化”病毒基因表达为蛋白质。
