Huang Zhihong, Pan Mengjia, Zhu Silei, Zhang Hao, Wu Wenbi, Yuan Meijin, Yang Kai
State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou, China.
State Key Laboratory of Biocontrol, Sun Yat-sen University, Guangzhou, China
J Virol. 2017 Feb 14;91(5). doi: 10.1128/JVI.02110-16. Print 2017 Mar 1.
is a family of insect-specific viruses that have a circular double-stranded DNA genome packaged within a rod-shaped capsid. The mechanism of baculovirus nucleocapsid assembly remains unclear. Previous studies have shown that deletion of the gene of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) blocks viral nucleocapsid assembly. Interestingly, the -encoded protein Ac83 is not a component of the nucleocapsid, implying a particular role for in nucleocapsid assembly that may be independent of its protein product. To examine this possibility, Ac83 synthesis was disrupted by insertion of a chloramphenicol resistance gene into its coding sequence or by deleting its promoter and translation start codon. Both mutants produced progeny viruses normally, indicating that the Ac83 protein is not required for nucleocapsid assembly. Subsequently, complementation assays showed that the production of progeny viruses required the presence of in the AcMNPV genome instead of its presence in Therefore, we reasoned that is involved in nucleocapsid assembly via an internal -acting element, which we named the nucleocapsid assembly-essential element (NAE). The NAE was identified to lie within nucleotides 1651 to 1850 of and had 8 conserved A/T-rich regions. Sequences homologous to the NAE were found only in alphabaculoviruses and have a conserved positional relationship with another essential -acting element that was recently identified. The identification of the NAE may help to connect the data of viral -acting elements and related proteins in the baculovirus nucleocapsid assembly, which is important for elucidating DNA-protein interaction events during this process. Virus nucleocapsid assembly usually requires specific -acting elements in the viral genome for various processes, such as the selection of the viral genome from the cellular nucleic acids, the cleavage of concatemeric viral genome replication intermediates, and the encapsidation of the viral genome into procapsids. In linear DNA viruses, such elements generally locate at the ends of the viral genome; however, most of these elements remain unidentified in circular DNA viruses (including baculovirus) due to their circular genomic conformation. Here, we identified a nucleocapsid assembly-essential element in the AcMNPV (the archetype of baculovirus) genome. This finding provides an important reference for studies of nucleocapsid assembly-related elements in baculoviruses and other circular DNA viruses. Moreover, as most of the previous studies of baculovirus nucleocapsid assembly have been focused on viral proteins, our study provides a novel entry point to investigate this mechanism via -acting elements in the viral genome.
杆状病毒是一类昆虫特异性病毒,其具有包装在杆状衣壳内的环状双链DNA基因组。杆状病毒核衣壳组装的机制仍不清楚。先前的研究表明,苜蓿银纹夜蛾多粒包埋核型多角体病毒(AcMNPV)的该基因缺失会阻断病毒核衣壳的组装。有趣的是,该基因编码的蛋白Ac83不是核衣壳的组成成分,这意味着该基因在核衣壳组装中可能具有独立于其蛋白产物的特殊作用。为了检验这种可能性,通过将氯霉素抗性基因插入其编码序列或删除其启动子和翻译起始密码子来破坏Ac83的合成。两种突变体均正常产生子代病毒,这表明核衣壳组装不需要Ac83蛋白。随后,互补试验表明,子代病毒的产生需要AcMNPV基因组中存在该基因,而不是其存在于……中。因此,我们推断该基因通过一个内部顺式作用元件参与核衣壳组装,我们将其命名为核衣壳组装必需元件(NAE)。NAE被确定位于该基因的1651至1850核苷酸内,并有8个保守的富含A/T的区域。与NAE同源的序列仅在α杆状病毒中发现,并且与最近鉴定的另一个必需顺式作用元件具有保守的位置关系。NAE的鉴定可能有助于将杆状病毒核衣壳组装中病毒顺式作用元件和相关蛋白的数据联系起来,这对于阐明该过程中的DNA-蛋白质相互作用事件很重要。病毒核衣壳组装通常需要病毒基因组中的特定顺式作用元件来进行各种过程,例如从细胞核酸中选择病毒基因组、切割串联病毒基因组复制中间体以及将病毒基因组包装到原衣壳中。在线状DNA病毒中,这些元件通常位于病毒基因组的末端;然而,由于其环状基因组构象,这些元件中的大多数在环状DNA病毒(包括杆状病毒)中仍未被鉴定。在这里,我们在AcMNPV(杆状病毒的原型)基因组中鉴定了一个核衣壳组装必需元件。这一发现为杆状病毒和其他环状DNA病毒中核衣壳组装相关元件的研究提供了重要参考。此外,由于先前对杆状病毒核衣壳组装的大多数研究都集中在病毒蛋白上,我们的研究通过病毒基因组中的顺式作用元件为研究这一机制提供了一个新的切入点。