Nuclear import of moloney murine leukemia virus DNA mediated by adenovirus preterminal protein is not sufficient for efficient retroviral transduction in nondividing cells.
Nuclear import of moloney murine leukemia virus DNA mediated by adenovirus preterminal protein is not sufficient for efficient retroviral transduction in nondividing cells.
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由腺病毒前末端蛋白介导的莫洛尼鼠白血病病毒DNA的核输入不足以在非分裂细胞中进行有效的逆转录病毒转导。
DOI：
10.1128/jvi.74.2.721-734.2000
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发表时间：
2000
影响因子：
5.4
通讯作者：
Li,ZY
中科院分区：
文献类型：
--
作者：
Lieber,A;Kay,MA;Li,ZY
Moloney小鼠白血病病毒（MoMLV）衍生载体需要细胞分裂才能有效转导，这可能与病毒dna -蛋白复合物无法穿过核膜有关。相反，腺病毒（Ad）可以有效地感染非分裂细胞。这种特性可能是由于许多Ad蛋白中存在多个核易位信号，这些信号与传入的病毒基因组相关。特别令人感兴趣的是Ad前端蛋白（pTP），它单独或与Ad聚合酶复合结合到Ad反向末端重复序列中的特定序列。本研究的目的是测试pTP与携带pTP结合位点的逆转录病毒DNA的共表达是否会促进病毒前整合复合体的核输入和静止细胞的转导。在初步实验中，我们证明了核pTP可以将质粒DNA共同导入生长受阻细胞的细胞核中。逆转录病毒转导研究是用G1/ s阻滞的LTA细胞或静止期人原代成纤维细胞进行的。这些研究表明，当逆转录病毒载体含有相应的结合基序时，pTP或pTP- ad聚合酶将逆转录病毒DNA的细胞核输入阻滞细胞。然而，ptp介导的MoMLV DNA在非分裂细胞中的核易位不足以实现稳定的转导。在S期或DNA修复合成期间激活的其他细胞因子需要有效的逆转录病毒整合。
Moloney murine leukemia virus (MoMLV)-derived vectors require cell division for efficient transduction, which may be related to an inability of the viral DNA-protein complex to cross the nuclear membrane. In contrast, adenoviruses (Ad) can efficiently infect nondividing cells. This property may be due to the presence of multiple nuclear translocation signals in a number of Ad proteins, which are associated with the incoming viral genomes. Of particular interest is the Ad preterminal protein (pTP), which binds alone or in complex with the Ad polymerase to specific sequences in the Ad inverted terminal repeat. The goal of this study was to test whether coexpression of pTP with retroviral DNA carrying pTP-binding sites would facilitate nuclear import of the viral preintegration complex and transduction of quiescent cells. In preliminary experiments, we demonstrated that the karyophylic pTP can coimport plasmid DNA into the nuclei of growth-arrested cells. Retroviral transduction studies were performed with G1/S-arrested LTA cells or stationary-phase human primary fibroblasts. These studies demonstrated that pTP or pTP-Ad polymerase conferred nuclear import of retroviral DNA upon arrested cells when the retrovirus vector contained the corresponding binding motifs. However, pTP-mediated nuclear translocation of MoMLV DNA in nondividing cells was not sufficient for stable transduction. Additional cellular factors activated during S phase or DNA repair synthesis were required for efficient retroviral integration.