Rescue of dnapol-null Autographa californica multiple nucleopolyhedrovirus with DNA polymerase (DNApol) of Spodoptera litura nucleopolyhedrovirus (SpltNPV) and identification of a nuclear localization signal in SpltNPV DNApol

文献类型： 外文期刊

第一作者： Chen, Guoqing

作者： Chen, Guoqing;Fang, Yang;Feng, Guozhong;Fang, Yang;Hu, Zhaoyang;Krell, Peter J.

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期刊名称：JOURNAL OF GENERAL VIROLOGY （ 影响因子：3.891； 五年影响因子：3.719 ）

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收录情况： SCI

摘要： DNA polymerase (DNApol) is highly conserved in all baculoviruses and plays an essential role in viral DNA replication. It determines the fidelity of baculovirus DNA replication by inserting the correct nucleotides into the primer terminus and proofreading any mispaired nucleotides. DNApols of groups I and II of the genus Alphabaculovirus in the family Baculoviridae share many common structural features. However, it is not clear whether a group I Autographa californica multiple nucleopolyhedrovirus (AcMNPV) DNApol can be substituted by a group II NPV DNApol. Here we report the successful generation of AcMNPV dnapol-null virus being rescued by a group II Spodoptera litura NPV (SpltNPV) dnapol (Bac-Delta cDPol : Slpol). Viral growth curves and quantitative real-time PCR showed that the dnapol replacement reduced the level of viral production and DNA replication of Bac-Delta cDPol : SlPol compared with WTrep, a native dnapol insertion in an AcMNPV dnapol-null virus. Light microscopy showed that production of occlusion bodies for Bac-Ac Delta Pol : Slpol was reduced. We also identified a nuclear localization signal (NLS) for the SpltNPV DNApol C terminus at residues 827-838 by mutational analysis and confocal microscopy. Multiple point substitution of SpltNPV DNApol NLS abrogated virus production and viral DNA replication. Overall, these data suggested that the NLS plays an important role in SpltNPV DNApol nuclear localization and that SpltNPV DNApol cannot efficiently substitute the AcMNPV DNApol in AcMNPV.

分类号： R37

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