pUL34 binding near the human cytomegalovirus origin of lytic replication enhances DNA replication and viral growth.

pUL34 binding near the human cytomegalovirus origin of lytic replication enhances DNA replication and viral growth.
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DOI:
10.1016/j.virol.2018.03.017
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发表时间:
2018-05
期刊:
影响因子:
3.7
通讯作者:
Biegalke BJ
Biegalke BJ
中科院分区:
医学3区
文献类型:
--
作者:
Slayton M;Hossain T;Biegalke BJ

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人巨细胞病毒(HCMV)UL 34基因编码病毒复制所需的序列特异性DNA结合蛋白(pUL 34)。pUL 34与DNA结合位点的相互作用抑制两个病毒免疫逃避基因US 3和US 9的转录。在HCMV基因组(菌株AD 169)中存在12个额外的预测pUL 34结合位点,其中3个结合位点集中在HCMV裂解复制起点(oriLyt)附近。我们使用pUL 34-DNA相互作用的ChIP-seq分析来确认pUL 34在感染期间结合oriLyt区域。在含有oriLy的质粒中的UL 34结合位点的突变显著降低了病毒介导的oriLy依赖性DNA复制。HCMV基因组中这些位点的突变降低了所得病毒的复制效率。蛋白质-蛋白质相互作用分析表明,pUL 34与病毒蛋白IE 2,UL 44和UL 84相互作用,这是病毒DNA复制所必需的,表明在oriLyt区域中的pUL 34-DNA相互作用参与DNA复制级联。
The human cytomegalovirus (HCMV) UL34 gene encodes sequence-specific DNA-binding proteins (pUL34) which are required for viral replication. Interactions of pUL34 with DNA binding sites represses transcription of two viral immune evasion genes, US3 and US9. 12 additional predicted pUL34-binding sites are present in the HCMV genome (strain AD169) with three binding sites concentrated near the HCMV origin of lytic replication (oriLyt). We used ChIP-seq analysis of pUL34-DNA interactions to confirm that pUL34 binds to the oriLyt region during infection. Mutagenesis of the UL34-binding sites in an oriLyt-containing plasmid significantly reduced viral-mediated oriLyt-dependent DNA replication. Mutagenesis of these sites in the HCMV genome reduced the replication efficiencies of the resulting viruses. Protein-protein interaction analyses demonstrated that pUL34 interacts with the viral proteins IE2, UL44, and UL84, that are essential for viral DNA replication, suggesting that pUL34-DNA interactions in the oriLyt region are involved in the DNA replication cascade.
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