Dong Zhan-Qi, Hu Nan, Zhang Jun, Chen Ting-Ting, Cao Ming-Ya, Li Hai-Qing, Lei Xue-Jiao, Chen Peng, Lu Cheng, Pan Min-Hui
State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, 400716, China.
Institutes of Life Sciences, Chongqing Medical University, Chongqing, 400716, China.
PLoS One. 2015 Dec 14;10(12):e0144930. doi: 10.1371/journal.pone.0144930. eCollection 2015.
We have previously reported that baculovirus Bombyx mori nucleopolyhedrovirus (BmNPV) late expression factor 11 (lef-11) is associated with viral DNA replication and have demonstrated that it potentially interacts with itself; however, whether LEF-11 forms oligomers and the impact of LEF-11 oligomerization on viral function have not been substantiated. In this study, we first demonstrated that LEF-11 is capable of forming oligomers. Additionally, a series of analyses using BmNPV LEF-11 truncation mutants indicated that two distinct domains control LEF-11 oligomerization (aa 42-61 and aa 72-101). LEF-11 truncation constructs were inserted into a lef-11-knockout BmNPV bacmid, which was used to demonstrate that truncated LEF-11 lacking either oligomerization domain abrogates viral DNA replication. Finally, site-directed mutagenesis was used to determine that the conserved hydrophobic residues Y58&I59 (representing Y58 and I59), I85 and L88&L89 (representing L88 and L89) are required for LEF-11 oligomerization and viral DNA replication. Collectively, these data indicate that BmNPV LEF-11 oligomerization influences viral DNA replication.
我们之前报道过,杆状病毒家蚕核型多角体病毒(BmNPV)晚期表达因子11(lef-11)与病毒DNA复制相关,并且已经证明它可能与自身相互作用;然而,LEF-11是否形成寡聚体以及LEF-11寡聚化对病毒功能的影响尚未得到证实。在本研究中,我们首先证明了LEF-11能够形成寡聚体。此外,一系列使用BmNPV LEF-11截短突变体的分析表明,两个不同的结构域控制LEF-11寡聚化(氨基酸42-61和氨基酸72-101)。将LEF-11截短构建体插入lef-11敲除的BmNPV杆粒中,用于证明缺乏任何一个寡聚化结构域的截短LEF-11会废除病毒DNA复制。最后,使用定点诱变来确定保守的疏水残基Y58&I59(代表Y58和I59)、I85以及L88&L89(代表L88和L89)是LEF-11寡聚化和病毒DNA复制所必需的。总的来说,这些数据表明BmNPV LEF-11寡聚化影响病毒DNA复制。