Carbone Mariarosaria, Reale Anna, Di Sauro Annarita, Sthandier Olga, Garcia Marie-Isabelle, Maione Rossella, Caiafa Paola, Amati Paolo
Pasteur Institute-Fondazione Cenci Bolognetti, Department of Cellular Biotechnology and Hematology, University of Rome La Sapienza, Viale Regina Elena 324, 00161 Rome, Italy.
J Mol Biol. 2006 Nov 3;363(4):773-85. doi: 10.1016/j.jmb.2006.05.077. Epub 2006 Jun 30.
Poly(ADP-ribose)polymerases are involved in fundamental cellular events as well as they seem to be associated to some viral infection process. In this work, the poly(ADP-ribose)polymerase-1 (PARP-1) role in the polyomavirus life cycle has been investigated. Early viral transcription was reduced by competitive inhibitors of PARPs in Swiss 3T3 cells and almost abolished in PARP-1 knockout fibroblasts and in wild-type fibroblasts when PARP-1 was silenced by RNA interference. In vivo chromatin immunoprecipitation assays showed that poly(ADP-ribosyl)ation (poly(ADP-ribose)) facilitates the release of the capsid protein viral protein 1 (VP1) from the chromatin of infecting virions. In vitro experiments demonstrated that VP1 stimulates the enzymatic activity of PARP-1 and binds non-covalently both protein-free and PARP-1-bound poly(ADP-ribose). Our studies suggest that PARP-1 promotes the complete VP1 displacement from viral DNA favouring the viral early transcription.
聚（ADP-核糖）聚合酶参与基本的细胞活动，而且似乎与某些病毒感染过程有关。在这项研究中，研究了聚（ADP-核糖）聚合酶-1（PARP-1）在多瘤病毒生命周期中的作用。在瑞士3T3细胞中，PARP的竞争性抑制剂可降低病毒早期转录，而在PARP-1基因敲除的成纤维细胞中以及在通过RNA干扰使PARP-1沉默的野生型成纤维细胞中，病毒早期转录几乎完全被消除。体内染色质免疫沉淀试验表明，聚（ADP-核糖基）化（聚（ADP-核糖））有助于衣壳蛋白病毒蛋白1（VP1）从感染性病毒粒子的染色质中释放出来。体外实验表明，VP1刺激PARP-1的酶活性，并与无蛋白的和PARP-1结合的聚（ADP-核糖）非共价结合。我们的研究表明，PARP-1促进VP1从病毒DNA上完全解离，从而有利于病毒早期转录。
