Shadan F F, Cowsert L M, Villarreal L P
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717.
J Virol. 1994 Aug;68(8):4785-96. doi: 10.1128/JVI.68.8.4785-4796.1994.
Small DNA viruses are dependent on the interaction of early proteins (such as large T antigen) with host p53 and Rb to bring about the G1-to-S cell cycle transition. The large DNA viruses are less dependent on host regulatory genes since additional early viral proteins (such as viral DNA polymerase, DNA metabolic enzymes, and other replication proteins) are involved in DNA synthesis. A highly conserved domain of large T antigen (similar to the p53-binding region) exclusively identifies papovavirus, parvovirus, and papillomaviruses from all other larger DNA viruses and implies a conserved interaction with host regulatory genes. In this report, we show that 3 to 6 mM butyrate, a general cell cycle blocker implicated in inhibition of the G1-to-S transition, inhibits DNA replication of polyomavirus and human papillomavirus type 11 but not the replication of larger DNA viruses such as adenovirus types 2 and 5, herpes simplex virus type 1, Epstein-Barr virus, and cytomegalovirus, which all bypass the butyrate-mediated cell cycle block. This butyrate effect on polyomavirus replication is not cell type specific, nor does it depend on the p53 or Rb gene, as inhibition was seen in fibroblasts with intact or homozygous deleted p53 or Rb, 3T6 cells, keratinocytes, C2C12 myoblasts, and 3T3-L1 adipocytes. In addition, butyrate did not inhibit expression of polyomavirus T antigen. The antiviral effect of butyrate involves a form of imprinted state, since pretreatment of cells with 3 mM butyrate inhibits human papillomavirus type 11 DNA replication for at least 96 h after its removal. Butyrate, therefore, serves as a molecular tool in dissecting the life cycle of smaller DNA viruses from that of the larger DNA viruses in relation to the cell cycle.
小型DNA病毒依赖于早期蛋白(如大T抗原)与宿主p53和Rb的相互作用来实现G1期到S期的细胞周期转变。大型DNA病毒对宿主调控基因的依赖性较小,因为额外的早期病毒蛋白(如病毒DNA聚合酶、DNA代谢酶和其他复制蛋白)参与DNA合成。大T抗原的一个高度保守结构域(类似于p53结合区域)专门识别乳头多瘤空泡病毒、细小病毒和乳头瘤病毒与所有其他更大的DNA病毒,这意味着与宿主调控基因存在保守的相互作用。在本报告中,我们表明3至6 mM丁酸盐是一种普遍的细胞周期阻滞剂,与抑制G1期到S期转变有关,它抑制多瘤病毒和人乳头瘤病毒11型的DNA复制,但不抑制更大的DNA病毒如腺病毒2型和5型、单纯疱疹病毒1型(HSV-1)、EB病毒和巨细胞病毒的复制,这些病毒都能绕过丁酸盐介导的细胞周期阻滞。丁酸盐对多瘤病毒复制的这种作用不是细胞类型特异性的,也不依赖于p53或Rb基因,因为在p53或Rb完整或纯合缺失的成纤维细胞、3T6细胞、角质形成细胞、C2C12成肌细胞和3T3-L1脂肪细胞中都观察到了抑制作用。此外,丁酸盐不抑制多瘤病毒T抗原的表达。丁酸盐的抗病毒作用涉及一种印记状态,因为用3 mM丁酸盐预处理细胞后,在其去除后至少96小时内抑制人乳头瘤病毒11型DNA复制。因此,丁酸盐作为一种分子工具,可用于剖析小型DNA病毒与大型DNA病毒在细胞周期方面的生命周期。