Epstein-Barr virus nuclear antigen 1 interacts with regulator of chromosome condensation 1 dynamically throughout the cell cycle
Epstein-Barr virus nuclear antigen 1 interacts with regulator of chromosome condensation 1 dynamically throughout the cell cycle
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DOI：
10.1099/jgv.0.000681
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发表时间：
2017-02-01
影响因子：
3.8
通讯作者：
Manet, Evelyne
中科院分区：
文献类型：
--
作者：
Deschamps, Thibaut;Bazot, Quentin;Manet, Evelyne
EB病毒（EBV）核抗原1（EBNA 1）是一种序列特异性DNA结合蛋白，其通过将DNA复制的细胞复合物募集到起点（oriP）上并通过将病毒DNA拴系到有丝分裂染色体上，在病毒附加体复制和分离中起重要作用。尽管病毒DNA复制的机制已经有了很好的记录，但那些涉及将EBNA 1束缚在细胞染色质上的机制还远未被理解。在这里，我们已经确定了染色体凝聚调节因子1（RCC 1）作为EBNA 1的一种新的细胞伴侣。RCC 1是小GTdR Ran酶的主要核鸟嘌呤核苷酸交换因子。与染色质相关的RCC 1参与RanGTP梯度的形成，RanGTP梯度对核质转运、有丝分裂纺锤体形成和有丝分裂后核膜重组至关重要。使用几种方法，我们已经证明了这两种蛋白质之间的直接相互作用，并发现负责EBNA 1拴系到有丝分裂染色体的EBNA 1结构域也参与了与RCC 1的相互作用。EBNA 1肽阵列的使用证实了RCC 1与这些区域的相互作用以及RCC 1的N-末端区域在这种相互作用中的重要性。最后，使用共聚焦显微镜和福斯特共振能量转移分析，以遵循整个细胞周期的两种蛋白质之间的相互作用的动力学，我们已经证明，EBNA 1和RCC 1密切相关的染色体在中期，这表明一个重要的作用，在这个阶段的相互作用，也许在拴EBNA 1有丝分裂染色体。
The Epstein-Barr virus (EBV) nuclear antigen 1 (EBNA1) is a sequence-specific DNA-binding protein that plays an essential role in viral episome replication and segregation, by recruiting the cellular complex of DNA replication onto the origin (oriP) and by tethering the viral DNA onto the mitotic chromosomes. Whereas the mechanisms of viral DNA replication are well documented, those involved in tethering EBNA1 to the cellular chromatin are far from being understood. Here, we have identified regulator of chromosome condensation 1 (RCC1) as a novel cellular partner for EBNA1. RCC1 is the major nuclear guanine nucleotide exchange factor for the small GTPase Ran enzyme. RCC1, associated with chromatin, is involved in the formation of RanGTP gradients critical for nucleo-cytoplasmic transport, mitotic spindle formation and nuclear envelope reassembly following mitosis. Using several approaches, we have demonstrated a direct interaction between these two proteins and found that the EBNA1 domains responsible for EBNA1 tethering to the mitotic chromosomes are also involved in the interaction with RCC1. The use of an EBNA1 peptide array confirmed the interaction of RCC1 with these regions and also the importance of the N-terminal region of RCC1 in this interaction. Finally, using confocal microscopy and Forster resonance energy transfer analysis to follow the dynamics of interaction between the two proteins throughout the cell cycle, we have demonstrated that EBNA1 and RCC1 closely associate on the chromosomes during metaphase, suggesting an essential role for the interaction during this phase, perhaps in tethering EBNA1 to mitotic chromosomes.