CELLULAR MUTATION MEDIATES T-ANTIGEN-POSITIVE REVERTANT CELLS RESISTANT TO SIMIAN VIRUS-40 TRANSFORMATION BUT NOT TO RETRANSFORMATION BY POLYOMAVIRUS AND ADENOVIRUS TYPE-2
CELLULAR MUTATION MEDIATES T-ANTIGEN-POSITIVE REVERTANT CELLS RESISTANT TO SIMIAN VIRUS-40 TRANSFORMATION BUT NOT TO RETRANSFORMATION BY POLYOMAVIRUS AND ADENOVIRUS TYPE-2
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DOI：
10.1128/jvi.61.6.1821-1827.1987
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发表时间：
1987-06-01
影响因子：
5.4
通讯作者：
GRAESSMANN, A
中科院分区：
文献类型：
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作者：
BAUER, M;GUHL, E;GRAESSMANN, A
用蛋氨酸饥饿法从全猴病毒40(SV40)转化的Fisher大鼠胚胎成纤维细胞(REF 52细胞)中分离出T抗原阳性转化回变株。转化细胞(SV-52细胞)的逆转是由细胞基因组内的突变引起的。为了证明这一点，我们从宿主基因组中提取了SV40 DNA，将其插入到pSPT18 DNA载体中，克隆到大肠杆菌中，并显微注射到REF 52细胞的细胞核中。获得了与显微注射野生型SV40 DNA相同的转化效率(20%~25%)的细胞，而且该细胞对SV40的再转化具有抵抗力。显微注射野生型SV40DNA I后，分离出42个独立的细胞系。所有分析品系的细胞都获得了额外的SV40DNA拷贝，但细胞形态或生长特性的变化不明显。然而，在多瘤病毒和腺病毒2型感染或显微注射后，回复突变体可以高效地再转化。此外，突变细胞与祖代REF 52细胞的融合导致了转化状态的恢复。
T-antigen-positive transformation revertant cell lines were isolated from fully simian virus 40 (SV40)-transformed Fisher rat embryo fibroblast cells (REF 52 cells) by methionine starvation. Reversion of the transformed cells (SV-52 cells) was caused by a mutation within the cellular genome. To demonstrate this, we isolated SV40 DNA from the host genome, inserted it into plasmid pSPT18 DNA, cloned it in Escherichia coli, and microinjected it into the nuclei of the REF 52 cells. Fully transformed cells were obtained with the same efficiency (20 to 25%) as after microinjection of wild-type SV40 DNA I. Furthermore, the revertant cells were resistant to retransformation by SV40. Following microinjection of wild-type SV40 DNA I, 42 independent cell lines were isolated. Cells of all analyzed lines acquired additional SV40 DNA copies, but changes in the cell morphology or growth characteristic were not demonstrable. However, the revertants were retransformable with a high efficiency after polyomavirus and adenovirus type 2 infections or microinjection. Also, fusion of the revertant cells with the grandparental REF 52 cells led to restoration of the transformed state.