Bukrinskaya A G, Miller G G, Lebedeva E N, Agaphonova L V, Masurenko N N, Ilyin K V
J Virol. 1974 Oct;14(4):924-33. doi: 10.1128/JVI.14.4.924-933.1974.
A particles with the diameter of 70 to 80 nm were isolated from the cytoplasm of HEp-2, HeLa, and AO cells producing oncornavirus of Mason-Pfizer-like type. Most of the A particles banded at 1.23 to 1.24 g/ml, whereas 3 to 10% banded at 1.29 g/ml in equilibrium sucrose gradients. They banded at 1.30 g/ml in CsCl gradients suggesting that they contained 8% RNA. Individual A particles sedimented at 200 to 250S in velocity sucrose gradients, but their significant part was found aggregated and sedimented at more than 300S. They were resistant to RNase digestion. A particles possessed polymerase activity which was preferentially activated by Mn(2+) rather than by Mg(2+), the RNA template being 60S RNA. Cross-hybridization with two DNA products and immunoassay showed that A particles and Mason-Pfizer-like oncornavirus produced by the same cells contained neither homological RNA sequences nor common antigens, suggesting that A particles are not intracellular precursors of Mason-Pfizer-like oncornavirus but represent an independent oncornavirus. Hybridization of A particle RNA with excess of cellular DNA revealed about 20 proviral copies per HEp-2 cell genome and no proviral copies in human embryo and placenta cell genomes.
从产生梅森 - 辉瑞样肿瘤病毒的HEp - 2、HeLa和AO细胞的细胞质中分离出直径为70至80纳米的A颗粒。在平衡蔗糖梯度中,大多数A颗粒的密度带为1.23至1.24克/毫升,而3%至10%的密度带为1.29克/毫升。在氯化铯梯度中它们的密度带为1.30克/毫升,表明它们含有8%的RNA。在速度蔗糖梯度中,单个A颗粒的沉降系数为200至250S,但发现它们的大部分聚集且沉降系数超过300S。它们对核糖核酸酶消化具有抗性。A颗粒具有聚合酶活性,该活性优先被锰离子而非镁离子激活,RNA模板为60S RNA。与两种DNA产物的交叉杂交和免疫测定表明,由相同细胞产生的A颗粒和梅森 - 辉瑞样肿瘤病毒既不含有同源RNA序列也不含有共同抗原,这表明A颗粒不是梅森 - 辉瑞样肿瘤病毒的细胞内前体,而是代表一种独立的肿瘤病毒。A颗粒RNA与过量细胞DNA的杂交显示,每个HEp - 2细胞基因组约有20个前病毒拷贝,而在人类胚胎和胎盘细胞基因组中没有前病毒拷贝。