The 32 kDa subunit of replication protein A (RPA) participates in the DNA replication of Mung bean yellow mosaic India virus (MYMIV) by interacting with the viral Rep protein.
The 32 kDa subunit of replication protein A (RPA) participates in the DNA replication of Mung bean yellow mosaic India virus (MYMIV) by interacting with the viral Rep protein.
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DOI：
10.1093/nar/gkl1088
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发表时间：
2007
影响因子：
14.9
通讯作者：
Mukherjee, Sunil Kumar
中科院分区：
文献类型：
--
作者：
Singh, Dharmendra Kumar;Islam, Mohammad Nurul;Choudhury, Nirupam Roy;Karjee, Sumona;Mukherjee, Sunil Kumar
绿豆黄花叶印度病毒（MYMIV）是begomoviridae属的成员，其基因组由DNA- a和DNA- b两部分组成，单链环状DNA约2.7 kb。在DNA滚圈复制（RCR）过程中，基因组的稳定性和复制起点茎环结构的维持至关重要。因此，研究了宿主单链dna结合蛋白复制蛋白A （RPA）在MYMIV RCR中的作用。从豌豆中分离出两个RPA亚基，即RPA70 kDa和RPA32 kDa，并在酵母系统中验证了它们的作用，该系统模拟了MYMIV DNA复制。本研究表明，在体外和酵母双杂交系统中，只有RPA32 kDa亚基直接与MYMIV-Rep的羧基端相互作用。RPA32通过增强其atp酶，下调其缺口和关闭活性来调节Rep的功能。讨论了这些调节在病毒DNA复制过程中的可能作用。最后，我们利用基于植物的实验证明了RPA32在携带MYMIV复制起源的质粒DNA的瞬时复制中的积极参与。
Mung bean yellow mosaic India virus (MYMIV) is a member of genus begomoviridae and its genome comprises of bipartite (two components, namely DNA-A and DNA-B), single-stranded, circular DNA of about 2.7 kb. During rolling circle replication (RCR) of the DNA, the stability of the genome and maintenance of the stem–loop structure of the replication origin is crucial. Hence the role of host single-stranded DNA-binding protein, Replication protein A (RPA), in the RCR of MYMIV was examined. Two RPA subunits, namely the RPA70 kDa and RPA32 kDa, were isolated from pea and their roles were validated in a yeast system in which MYMIV DNA replication has been modelled. Here, we present evidences that only the RPA32 kDa subunit directly interacted with the carboxy terminus of MYMIV-Rep both in vitro as well as in yeast two-hybrid system. RPA32 modulated the functions of Rep by enhancing its ATPase and down regulating its nicking and closing activities. The possible role of these modulations in the context of viral DNA replication has been discussed. Finally, we showed the positive involvement of RPA32 in transient replication of the plasmid DNA bearing MYMIV replication origin using an in planta based assay.
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