PCNA interacts with Indian mung bean yellow mosaic virus rep and downregulates rep activity
PCNA interacts with Indian mung bean yellow mosaic virus rep and downregulates rep activity
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DOI：
10.1128/jvi.78.21.11890-11903.2004
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发表时间：
2004-11-01
影响因子：
5.4
通讯作者：
Mukherjee, SK
中科院分区：
文献类型：
--
作者：
Bagewadi, B;Chen, SJ;Mukherjee, SK
增殖细胞核抗原（PCNA）是一种保守的植物蛋白，也是一种重要的复制因子，在双生病毒感染的韧皮部组织的静息细胞中被诱导。PCNA在双生病毒DNA滚环复制（RCR）中的生物化学作用尚未详细研究。印度绿豆黄花叶病毒（Indian mung bean yellow mosaic virus，IMYMV）是一种双生病毒，其二分基因组的RCR启动主要受病毒蛋白Rep（或AL 1或AC 1）控制。通过研究重组PCNA和重组IMYMV Rep. Pea核PCNA以及重组豌豆PCNA之间的物理和功能相互作用，揭示了宿主PCNA在IMYMV RCR中的作用。在涉及亲和层析和酵母双杂交方法的实验中，重组豌豆PCNA与重组Rep结合。PCNA的接触氨基酸残基似乎存在于整个三聚体蛋白的广泛区域，而Rep的那些似乎仅定位于蛋白的中间部分。PCNA可使IMYMV Rep的位点特异性切口闭合活性和ATP酶功能受损。这些观察结果导致有趣的推测控制病毒RCR和蛋白质-蛋白质相互作用的动态配置文件在RCR起源的双生病毒。
Proliferative cell nuclear antigen (PCNA), a conserved plant protein as well as an important replication factor, is induced in response to geminivirus infection in the resting cells of the phloem tissues. The biochemical role of PCNA in rolling circle replication (RCR) of geminivirus DNA has not been explored in detail. The initiation of RCR of the bipartite genome of a geminivirus, Indian mung bean yellow mosaic virus (IMYMV), is mainly controlled by viral protein Rep (or AL1 or AC1). The role of host PCNA in RCR of IMYMV was revealed by studying the physical and functional interactions between recombinant PCNA and recombinant IMYMV Rep. Pea nuclear PCNA as well as recombinant pea PCNA showed binding to recombinant Rep in experiments involving both affinity chromatography and yeast two-hybrid approaches. The contacting amino acid residues of PCNA seemed to be present throughout a wide region of the trimeric protein, while those of Rep appeared to be localized only in the middle part of the protein. The site-specific nicking-closing activity and the ATPase function of IMYMV Rep were impaired by PCNA. These observations lead to interesting speculations about the control of viral RCR and dynamic profiles of protein-protein interactions at the RCR origin of the geminiviruses.