Genome-wide chromatin interaction profiling reveals a vital role of super-enhancers and rearrangements in host enhancer contacts during BmNPV infection.

As influential regulatory elements in the genome, enhancers control gene expression under specific cellular conditions, and such connections are dynamic under different conditions. However, because of the lack of a genome-wide enhancer-gene connection map, the roles and regulatory pattern of enhancers were poorly investigated in insects, and the dynamic changes of enhancer contacts and functions under different conditions remain elusive. Here, combining Hi-C, ATAC-seq, and H3K27ac ChIP-seq data, we generate the genome-wide enhancer-gene map of silkworm and identify super-enhancers with a role in regulating the expression of vital genes related to cell state maintenance through a sophisticated interaction network. Additionally, a radical attenuation of chromatin interactions is found after infection of Bombyx mori nucleopolyhedrovirus (BmNPV), the main pathogen of silkworm, which directly rearranges the enhancer contacts. Such a rearrangement disturbs the intrinsic enhancer-gene connections in several antiviral genes, resulting in reduced expression of these genes, which accelerates viral infection. Overall, our results reveal the regulatory role of super-enhancers and shed new light on the mechanisms and dynamic changes of the genome-wide enhancer regulatory network.

Overexpression Bombyx mori HEXIM1 Facilitates Immune Escape of Bombyx mori Nucleopolyhedrovirus by Suppressing BmRelish-Driven Immune Responses.

Bombyx mori nucleopolyhedrovirus (BmNPV), a typical arthropod-specific enveloped DNA virus, is one of the most serious pathogens in silkworm farming, but the potential mechanisms of the evasion of innate immune responses from BmNPV infection are still poorly understood. HEXIM1 is an RNA-binding protein, best known as an inhibitor of positive transcription elongation factor b (P-TEFb), which controls transcription elongation by RNA polymerase II. In this study, Bombyx mori HEXIM1 (BmHEXIM1) was cloned and characterized, and its expression was found to be remarkably upregulated after BmNPV infection. Furthermore, BmHEXIM1 was detected to increase the proliferation of BmNPV, and its full length is essential for assisting BmNPV immune escape by suppressing BmRelish-driven immune responses. This study brought new insights into the mechanisms of immune escape of BmNPV and provided theoretical guidance for the breeding of BmNPV-resistant silkworm varieties.

BmNPV p35 Reduces the Accumulation of Virus-Derived siRNAs and Hinders the Function of siRNAs to Facilitate Viral Infection.

Antiviral immunity involves various mechanisms and responses, including the RNA interference (RNAi) pathway. During long-term coevolution, viruses have gained the ability to evade this defense by encoding viral suppressors of RNAi (VSRs). It was reported that p35 of baculovirus can inhibit cellular small interference RNA (siRNA) pathway; however, the molecular mechanisms underlying p35 as a VSR remain largely unclear. Here, we showed that p35 of Bombyx mori nucleopolyhedrovirus (BmNPV) reduces the accumulation of virus-derived siRNAs (vsiRNAs) mapped to a particular region in the viral genome, leading to an increased expression of the essential genes in this region, and revealed that p35 disrupts the function of siRNAs by preventing them from loading into Argonaute-2 (Ago2). This repressive effect on the cellular siRNA pathway enhances the replication of BmNPV. Thus, our findings illustrate for the first time the inhibitory mechanism of a baculovirus VSR and how this effect influences viral infection.

Molecular mechanism responsible for the hyperexpression of baculovirus polyhedrin.

One of the amazing phenomena in the baculovirus life cycle is the hyperexpression of the very late gene, polyhedrin (polh), causing the production of the occlusion bodies where progeny virions are embedded. However, to date, the molecular mechanism underlying its hyperexpression is not completely elucidated. Considering that, in this review, the mechanism responsible for its hyperexpression from the previous studies up to now was comprehensively summarized from three aspects, namely, the structure characteristics of the polh promoter and transcription regulation, the structure and translation regulation of the polh mRNA, and especially the regulators that influence the expression of polh gene. Moreover, this review will help us obtain a better understanding about the hyperexpression of polh, and also provide guidance for improving the expression efficiency of the foreign proteins by adopting the baculovirus expression vector system.

Genome-wide identification of lipases in silkworm (Bombyx mori) and their spatio-temporal expression in larval midgut.

Lipases play crucial roles in food digestion by degrading dietary lipids into free fatty acids and glycerols. The domesticated silkworm (Bombyx mori) has been widely used as an important Lepidopteran model for decades. However, little is known about the lipase gene family in the silkworm, especially their hydrolytic activities as digestive enzymes. In this study, a total of 38 lipase genes were identified in the silkworm genome. Phylogenetic analysis indicated that they were divided into three major groups. Twelve lipases were confirmed to be expressed in the midgut at both transcriptional and translational levels. They were grouped into the same gene cluster, suggesting that they could have similar physiological functions. Quantitative real-time PCR (qRT-PCR) analyses indicated that lipases were mainly expressed in anterior and middle midgut regions, and their expression levels varied greatly along the length of midgut. A majority of lipases were down-regulated in the midgut when larvae stopped feeding. However, a unique lipase gene (Bmlip10583) showed low expression level during feeding stage, but it was significantly up-regulated during the larvae-pupae transition. These results demonstrated that expression of silkworm lipases was spatially and temporally regulated in the midgut during larval development. Taken together, our results provide a fundamental research of the lipase gene family in the silkworm.

RNAi-based immunity in insects against baculoviruses and the strategies of baculoviruses involved in siRNA and miRNA pathways to weaken the defense.

Protection against viral infection in hosts concerns diverse cellular and molecular mechanisms, among which RNA interference (RNAi) response is a vital one. Small interfering RNAs (siRNAs), microRNAs (miRNAs) and PIWI interacting RNAs (piRNAs) are primary categories of small RNAs involved in RNAi response, playing significant roles in restraining viral invasion. However, during a long-term coevolution, viruses have gained the ability to evade, avoid, or suppress antiviral immunity to ensure efficient replication and transmission. Baculoviruses are enveloped, insect-pathogenic viruses with double-stranded circular DNA genomes, which encode suppressors of siRNA pathway and miRNAs targeting immune-related genes to mask the antiviral activity of their hosts. This review summarized recent findings for the RNAi-based antiviral immunity in insects as well as the strategies that baculoviruses exploit to break the shield of host siRNA pathway, and hijack cellular miRNAs or encode their own miRNAs that regulate both viral and cellular gene expression to create a favorable environment for viral infection.

BmNPV-induced hormone metabolic disorder in silkworm leads to enhanced locomotory behavior.

Many parasites alter the host locomotory behaviors in a way that increases their fitness and progeny transmission. Baculoviruses can manipulate host physiology and alter the locomotory behavior by inducing 'hyperactivity' (increased locomotion) or 'tree-top disease' (climbing high up to the top before dying). However, the detailed molecular mechanism underlying virus-induced this hyperactive behavior remains elusive. In the present study, we showed that BmNPV invaded into silkworm brain tissue, resulting in severe brain damage. Moreover, BmNPV infection disturbed the insect hormone balance. The content of 20-hydroxyecdysone (20E) in hemolymph was much lower during the hyperactive stage, while the dopamine (DA) titer was higher than mock infection. Exogenous hormone treatment assays demonstrated that 20E inhibits virus-induced ELA (enhanced locomotory activity), while dopamine stimulates this behavior. More specificity, injection of dopamine or its agonist promote this hyperactive behavior in BmNPV-infected larvae. Taking together, our findings revealed the important role of hormone metabolism in BmNPV-induced ELA.

Structure-dependent antimicrobial mechanism of quaternary ammonium resins and a novel synthesis of highly efficient antimicrobial resin.

The demand for powerful and multifunctional water-treatment materials and reagents is increasing, because we are facing worse raw water quality, various tolerant bacteria, and risky disinfection by-products (DBPs) in drinking water. Quaternary ammonium resins (QARs) are promising candidates for water disinfection and purification, but their limited bactericidal capacities are difficult to improve because of the lack of guidelines for enhancing antibacterial efficiency. Therefore, we first systematically studied the structure-dependent antimicrobial mechanism of QARs and found that the best resin skeleton is acrylic-type, the optimal bactericidal alkyl is hexyl or octyl, the most applicable sizes are 80-100 meshes, the best counter anion is iodide ion, and the optimum quaternization reagent is iodoalkane. Moreover, the antibacterial capacity was demonstrated to depend on surficial N+ groups, correlating with surficial N+ charge density (R2 of 0.98) but not with exchange capacity (R2 of 0.26), physical adsorption of resin skeleton, or electrostatic adsorption of N+ groups. Based on these principles, we synthesized a new resin, Ac-81, with a surficial antibacterial design, which simultaneously exhibited better antimicrobial efficiency (two orders of magnitude) as well as higher contaminant removal potential (61.92%) compared to the traditional Ac-8C antibacterial resin. Furthermore, the new resin showed remarkable broad-spectrum antibacterial effects against Gram-negative E. coli and P. aeruginosa and Gram-positive B. subtilis and S. aureus in simulated water and actual water. Simultaneously, water quality was significantly improved, with HCO3-, SO42-, TN, TP, and TOC reduced by 79-90%, >99%, 66-85%, >99%, and 22-26%, respectively. Ac-81 is characterized by facile reusability, high treatment capacity of 1500 bed volume, and good adaptability for treating actual water, providing a promising alternative for drinking-water disinfection and purification.

Identification of A functional region in Bombyx mori nucleopolyhedrovirus VP39 that is essential for nuclear actin polymerization.

Nuclear actin polymerization plays an indispensable role in the nuclear assembly of baculovirus nucleocapsid, but the underlying viral infection-mediated mechanism remains unclear. VP39 is the major protein in baculovirus capsid, which builds the skeleton of the capsid tubular structure. VP39 is suggested in previous studies to interact with cellular actin and mediate actin polymerization. However, it is unclear about the role of VP39 in mediating nuclear actin polymerization. Results in this study indicated that vp39 deletion abolished nuclear actin polymerization, which was recovered after vp39 repair, revealing the essential part of VP39 in nuclear actin polymerization. Furthermore, a series of mutants with vp39 deletions were constructed to analyze the important region responsible for nuclear actin polymerization. In addition, intracellular localization analysis demonstrated that the amino acids 192-286 in VP39 C-terminal are responsible for nuclear actin polymerization.

Dynamic chromatin accessibility profiling reveals changes in host genome organization in response to baculovirus infection.

DNA viruses can hijack and manipulate the host chromatin state to facilitate their infection. Multiple lines of evidences reveal that DNA virus infection results in the host chromatin relocation, yet there is little known about the effects of viral infection on the architecture of host chromatin. Here, a combination of epigenomic, transcriptomic and biochemical assays were conducted to investigate the temporal dynamics of chromatin accessibility in response to Bombyx mori nucleopolyhedrovirus (BmNPV) infection. The high-quality ATAC-seq data indicated that progressive chromatin remodeling took place following BmNPV infection. Viral infection resulted in a more open chromatin architecture, along with the marginalization of host genome and nucleosome disassembly. Moreover, our results revealed that chromatin accessibility in uninfected cells was regulated by euchromatic modifications, whereas the viral-induced highly accessible chromatin regions were originally associated with facultative heterochromatic modification. Overall, our findings illustrate for the first time the organization and accessibility of host chromatin in BmNPV-infected cells, which lay the foundation for future studies on epigenomic regulation mediated by DNA viruses.

Post-translational modification of baculovirus-encoded proteins.

Post-translational modifications (PTMs) are the chemical modifications of proteins after translation, and are very important to guarantee the proper biological functions of these proteins. Baculoviruses are pathogenic viruses that infect invertebrates and have large circular double-stranded DNA genomes. Many proteins encoded by baculoviruses have been reported to have PTMs, including phosphorylation, glycosylation, ubiquitination, acetylation, and etc. However, up to now no overview of this information has been produced. In this review, we have summarized the PTMs that have been reported in baculovirus. As a majority of the studies on baculovirus have focused on Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and Bombyx mori nucleopolyhedrovirus(BmNPV), this review is focused primarily on these viruses.