Infection of primary cells by adeno-associated virus type 2 results in a modulation of cell cycle-regulating proteins
Infection of primary cells by adeno-associated virus type 2 results in a modulation of cell cycle-regulating proteins
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DOI：
10.1128/jvi.71.8.6020-6027.1997
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发表时间：
1997-08-01
影响因子：
5.4
通讯作者：
zurHausen, H
中科院分区：
文献类型：
--
作者：
Hermanns, J;Schulze, A;zurHausen, H
已证实腺相关病毒(AAV)感染原代人细胞可导致细胞增殖下降和生长停滞，我们分析了这种现象的分子基础，观察到AAV2感染对参与哺乳动物细胞周期控制的几个因子有影响，特别是参与G(1)细胞周期检查点控制的所有pRb家族成员p107和p130都受到影响。感染后，细胞周期蛋白A和B1分别在转录水平和蛋白水平下调，而G(1)/S细胞周期蛋白D1和E未受影响。此外，细胞周期蛋白依赖性激酶CDK1和CDK2以及转录因子E2F-1的稳态水平降低。在所有已知的参与pRB家族蛋白磷酸化的因子中，只有CDK抑制剂p21(WAF1)对AAV2感染有反应。至少72小时后，p21(WAF1)基因以非依赖于P53的方式迅速上调，与p21(WAF1)蛋白水平升高一致，细胞周期蛋白E和细胞周期蛋白A依赖的激酶活性下降到低水平，E2F-p130-细胞周期蛋白-CDK2复合体被破坏。这些数据表明，AAV2感染对原代人成纤维细胞的主要作用可能是上调p21(WAF1)基因的表达，从而抑制PRB家族蛋白的磷酸化，从而使细胞周期停滞。
It has been demonstrated that infection of primary human cells with adeno-associated viruses (AAV) leads to a decrease in cellular proliferation and to growth arrest, We analyzed the molecular basis of this phenomenon and observed that infection with AAV type 2 (AAV2) had an effect on several factors engaged in the control of the mammalian cell cycle, In particular, all of the pRB family members, pRB, p107, and p130, which are involved in G(1) cell cycle checkpoint control, were affected. After infection, a shift from hyper- to hypophosphorylated forms was observed, Cyclins A and B1, which are required for G(1)/S transition and progression into mitosis, respectively, were downregulated at the transcriptional level as well as at the protein level, whereas the G(1) cyclins D1 and E remained unaffected, In addition, the steady-state levels of cyclin-dependent kinases CDK1 and CDK2 and of transcription factor E2F-1 were diminished, Of all the factors known to be involved in phosphorylation of pRB family proteins, only the CDK inhibitor p21(WAF1) exhibited a response to AAV2 infection, p21(WAF1) mRNA was quickly and progressively upregulated in a p53-independent manner over at least 72 h, Consistent with the increased p21(WAF1) protein levels, cyclin E- and cyclin A-dependent kinase activities declined to low levels and E2F-p130-cyclin-CDK2 complexes were disrupted, From these data, we conclude that the major effect of AAV2 infection on primary human fibroblasts appears to be upregulation of p21(WAF1) gene expression and thus cell cycle arrest by the suppression of pRB family protein phosphorylation.