Guizani I, Kieny M P, Lathe R, Clertant P
INSERM U273, Centre de Biochimie du CNRS, Campus Valrose, Nice, France.
Gene. 1988 Dec 15;73(1):163-73. doi: 10.1016/0378-1119(88)90322-8.
We previously reported that live recombinant vaccinia viruses (VV) encoding either the large T (LT) or middle T (MT) antigens of polyoma virus (PyV) were able to induce rejection of tumors caused by PyV-transformed cells [Lathe et al., Nature 326 (1987) 878-880]. Here we present evidence that PyV early proteins expressed by the recombinants retain the biochemical characteristics of their authentic counterparts despite the cytopathic effect of VV infection. VV-encoded LT is a nuclear phosphoprotein, with specific DNA binding, ATPase and nucleotide-binding activities. VV-expressed MT associates with cellular kinases, particularly with pp60c-src, by which it is phosphorylated in vitro. Expression levels of LT and MT reached 10(6) molecules per infected cell. The use of VV as a vector is encouraged by the high expression level obtained and because VV infection does not seem to prevent appropriate post-translational processing of proteins encoded by VV recombinants.
我们先前报道过，编码多瘤病毒（PyV）大T（LT）或中T（MT）抗原的重组痘苗病毒（VV）能够诱导由PyV转化细胞引起的肿瘤的排斥反应[Lathe等人，《自然》326（1987）878 - 880]。在此我们提供证据表明，尽管存在VV感染的细胞病变效应，但重组体表达的PyV早期蛋白仍保留其天然对应物的生化特性。VV编码的LT是一种核磷蛋白，具有特定的DNA结合、ATP酶和核苷酸结合活性。VV表达的MT与细胞激酶相关联，特别是与pp60c-src相关联，并在体外被其磷酸化。LT和MT的表达水平达到每个感染细胞10^6个分子。由于获得了高表达水平，并且VV感染似乎并不妨碍对VV重组体编码的蛋白质进行适当的翻译后加工，因此鼓励使用VV作为载体。
