Huberdeau D, Sylla B S, Herring-Gillam E, Bourgaux-Ramoisy D, Bourgaux P
Mol Cell Biol. 1985 Oct;5(10):2608-12. doi: 10.1128/mcb.5.10.2608-2612.1985.
The Cyp cell line consists of mouse cells transformed by a thermosensitive polyomavirus (Py) genome and routinely propagated at 39 degrees C. Cyp cells are readily induced to synthesize free Py DNA by being transferred to 33 degrees C. In one subclone (C12/a1/S48, or S48) of this line, such induction resulted in the intracellular accumulation of three discrete species of cyclic DNA, i.e., genomic Py DNA, RmI, and RmII. RmI and RmII are Py-mouse chimeras, each of which contains a distinct set of sequences originating from the site of integration. Conceivably, genomic Py DNA, RmI, and RmII could persist at 39 degrees C as free replicating plasmids or originate from distinct populations of cells in S48 cultures. The data indicated that all three species arise at 33 degrees C from a genetically homogeneous cell population in which neither RmI nor RmII replicates at 39 degrees C. Examination of the sequence at the viral-cellular junction unique to RmII indicated that this chimera is excised from the host chromosome through a recombination event involving a complex viral sequence and a simple cellular sequence. Therefore, RmII provides another example of precise recombination occurring between nonhomologous sequences in a mammalian cell, as already observed for RmI (B. S. Sylla, D. Huberdeau, D. Bourgaux-Ramoisy, and P. Bourgaux, Cell 37:661-667, 1984).
Cyp细胞系由经温度敏感型多瘤病毒(Py)基因组转化的小鼠细胞组成,通常在39℃下传代培养。将Cyp细胞转移至33℃时,它们很容易被诱导合成游离的Py DNA。在该细胞系的一个亚克隆(C12/a1/S48,或S48)中,这种诱导导致细胞内积累三种不同的环状DNA,即基因组Py DNA、RmI和RmII。RmI和RmII是Py-小鼠嵌合体,每个都包含一组源自整合位点的独特序列。可以想象,基因组Py DNA、RmI和RmII在39℃时可能作为游离复制质粒持续存在,或者源自S48培养物中不同的细胞群体。数据表明,这三种类型均在33℃时从基因同质的细胞群体中产生,其中RmI和RmII在39℃时均不复制。对RmII特有的病毒-细胞连接处的序列进行检查表明,这种嵌合体是通过涉及复杂病毒序列和简单细胞序列的重组事件从宿主染色体上切除的。因此,RmII提供了另一个哺乳动物细胞中非同源序列之间发生精确重组的例子,这已在RmI中观察到(B.S.西拉、D.于伯德奥、D.布尔戈-拉穆瓦西和P.布尔戈,《细胞》37:661-667,1984年)。