Goldman N, Brown M, Khoury G
Cell. 1981 May;24(2):567-72. doi: 10.1016/0092-8674(81)90347-0.
The SV40 large T antigen is a multifunctional protein presumed to represent a single translation product of the early viral genes. A wide range of biological controls including the regulation of viral DNA replication, transcription and cell transformation has been attributed to T antigen. Previous evidence has indicated that large T antigen is modified in at least two ways, N-terminal acetylation and amino acid phosphorylation. In this study, we demonstrate a novel modification of a population of SV40 T antigen molecules by poly ADP-ribosylation. The covalent linkage of this oligonucleotide side chain to large T antigen, but not small t antigen, was demonstrated in experiments in which SV40-infected cells were labeled in vivo with 32P-orthophosphate or 14C-adenosine. Treatment of this labeled T antigen with snake venom phosphodiesterase released iso-ADP-ribose or treatment with ADP-ribose glycohydrolase released ADP-ribose. A method has been developed for the in vitro ADP-ribosylation of T antigen present in an infected cell nuclear extract with radiolabeled NAD. Since this type of modification is known to affect enzyme activity, its presence on T antigen suggests a similar role in the regulation of certain biological functions under the control of this protein.
SV40大T抗原是一种多功能蛋白,被认为是早期病毒基因的单一翻译产物。包括病毒DNA复制、转录和细胞转化调控在内的多种生物学控制都归因于T抗原。先前的证据表明,大T抗原至少以两种方式被修饰,即N端乙酰化和氨基酸磷酸化。在本研究中,我们证明了一群SV40 T抗原分子通过多聚ADP-核糖基化发生了一种新的修饰。在用32P-正磷酸盐或14C-腺苷对SV40感染的细胞进行体内标记的实验中,证实了这种寡核苷酸侧链与大T抗原而非小t抗原的共价连接。用蛇毒磷酸二酯酶处理这种标记的T抗原可释放异ADP-核糖,或用ADP-核糖糖苷水解酶处理可释放ADP-核糖。已经开发出一种方法,用放射性标记的NAD对感染细胞核提取物中存在的T抗原进行体外ADP-核糖基化。由于已知这种修饰会影响酶活性,它在T抗原上的存在表明在该蛋白控制下的某些生物学功能的调节中可能起类似作用。