The chromatin remodeling factor SMARCB1 forms a complex with human cytomegalovirus proteins UL114 and UL44.
The chromatin remodeling factor SMARCB1 forms a complex with human cytomegalovirus proteins UL114 and UL44.
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DOI：
10.1371/journal.pone.0034119
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Bjørås M
中科院分区：
文献类型：
--
作者：
Ranneberg-Nilsen T;Rollag H;Slettebakk R;Backe PH;Olsen Ø;Luna L;Bjørås M
人巨细胞病毒（HCMV）尿嘧啶DNA糖基化酶UL114是病毒DNA高效复制所必需的。据推测，UL114作为DNA复制机制中其他因子的结构伴侣发挥作用，而非作为一种DNA修复蛋白。UL114与UL44（HCMV持续合成因子）和UL54（HCMV - DNA聚合酶）结合。在本研究中，我们寻找了UL114的细胞伴侣。
在酵母双杂交筛选中，发现SWI/SNF染色质重塑复合物的一个因子SMARCB1是UL114的相互作用伴侣。这种相互作用通过免疫共沉淀和下拉实验在体外得到证实。免疫荧光显微镜显示，SMARCB1以及BRG - 1、BAF170和BAF155（这些是有效染色质重塑所需的SWI/SNF核心成分）在感染后24 - 48小时（hpi）出现在病毒复制灶中。此外，还证明了SMARCB1和UL44之间存在直接相互作用。
有效染色质重塑所需的SWI/SNF核心因子在整个感染过程中都存在于HCMV复制灶中。蛋白质UL44和UL114与SMARCB1相互作用，并可能参与将SWI/SNF复合物招募到染色质化的病毒DNA上。因此，SWI/SNF染色质重塑复合物在复制灶中的存在以及它与UL114和UL44的关联可能意味着它参与了不同的DNA活动。
Human cytomegalovirus (HCMV) uracil DNA glycosylase, UL114, is required for efficient viral DNA replication. Presumably, UL114 functions as a structural partner to other factors of the DNA-replication machinery and not as a DNA repair protein. UL114 binds UL44 (HCMV processivity factor) and UL54 (HCMV-DNA-polymerase). In the present study we have searched for cellular partners of UL114. In a yeast two-hybrid screen SMARCB1, a factor of the SWI/SNF chromatin remodeling complex, was found to be an interacting partner of UL114. This interaction was confirmed in vitro by co-immunoprecipitation and pull-down. Immunofluorescence microscopy revealed that SMARCB1 along with BRG-1, BAF170 and BAF155, which are the core SWI/SNF components required for efficient chromatin remodeling, were present in virus replication foci 24–48 hours post infection (hpi). Furthermore a direct interaction was also demonstrated for SMARCB1 and UL44. The core SWI/SNF factors required for efficient chromatin remodeling are present in the HCMV replication foci throughout infection. The proteins UL44 and UL114 interact with SMARCB1 and may participate in the recruitment of the SWI/SNF complex to the chromatinized virus DNA. Thus, the presence of the SWI/SNF chromatin remodeling complex in replication foci and its association with UL114 and with UL44 might imply its involvement in different DNA transactions.
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