Anderson K P, Lie Y S, Low M A, Williams S R, Wurm F M, Dinowitz M
Department of Medicinal and Analytical Chemistry, Genentech Inc., South San Francisco, CA 94080.
Dev Biol Stand. 1991;75:123-32.
The presence of budding C-type and intracytoplasmic A-type particles in Chinese hamster ovary (CHO) cells is well documented. However, extensive screening has failed to detect any evidence of infectivity. To investigate the origin and expression of these particles, retrovirus-like sequences which are actively transcribed in CHO cells have been cloned and characterized. Two families of sequences related to intracisternal A-particle (IAP) genomes of mice and Syrian hamsters were identified in cytoplasmic RNA from CHO cells (CHO IAP family I and family II). None of the four clones which were sequenced exhibited intact gag, pol, or env open reading frames. Only IAP family II sequences were present in purified extracellular particles of CHO cells. Several cDNA sequences related to mammalian C-type retrovirus genomes were isolated and cloned from gradient-purified, extracellular particles of recombinant CHO cells. All were homologous to the conserved endonuclease domain of murine leukemia virus. Nucleotide sequence analysis of the largest cDNA revealed multiple interruptions of the endonuclease encoding reading frame providing one possible explanation for the non-infectious nature of the particles observed in CHO cells. Both types of retrovirus-like sequences identified in purified extracellular particles of CHO cells (CHO IAP family II and C-type) were present as conserved, moderately repetitive sequences in DNA of all CHO cell lines examined, as well as in DNA from a Chinese hamster liver. It is therefore likely that the extracellular retrovirus-like particles of CHO cells are the products of endogenous provirus elements present in the germline of Chinese hamsters.
在中国仓鼠卵巢(CHO)细胞中存在出芽的C型和胞质A型颗粒,这一点已有充分记录。然而,广泛的筛选未能检测到任何感染性证据。为了研究这些颗粒的起源和表达,已克隆并鉴定了在CHO细胞中活跃转录的逆转录病毒样序列。在CHO细胞的细胞质RNA中鉴定出了与小鼠和叙利亚仓鼠的胞内A颗粒(IAP)基因组相关的两个序列家族(CHO IAP家族I和家族II)。测序的四个克隆中没有一个显示出完整的gag、pol或env开放阅读框。只有IAP家族II序列存在于CHO细胞纯化的细胞外颗粒中。从重组CHO细胞的梯度纯化细胞外颗粒中分离并克隆了几个与哺乳动物C型逆转录病毒基因组相关的cDNA序列。所有这些序列都与鼠白血病病毒的保守内切酶结构域同源。对最大的cDNA进行核苷酸序列分析发现,内切酶编码阅读框存在多处中断,这为在CHO细胞中观察到的颗粒的非感染性本质提供了一种可能的解释。在CHO细胞纯化的细胞外颗粒中鉴定出的两种逆转录病毒样序列(CHO IAP家族II和C型),在所有检测的CHO细胞系的DNA以及中国仓鼠肝脏的DNA中均作为保守的中度重复序列存在。因此,CHO细胞的细胞外逆转录病毒样颗粒很可能是中国仓鼠种系中存在的内源性前病毒元件的产物。