ABSTRACT Circular RNAs (circRNAs) are a class of endogenous non-coding RNAs widely expressed across diverse organisms. These molecules are involved in various biological processes, such as transcriptional and post-transcriptional regulation, protein scaffolding, and acting as miRNA sponges. Newcastle disease virus (NDV) replicates and proliferates in a variety of cells, inducing severe organelle stress damage, autophagy, and even cell death. Previous studies have reported that NDV hijacks and uses autophagy to promote its replication and proliferation. However, the involvement and regulatory mechanisms of circRNAs in this process remain largely unexplored. The present study aimed to identify novel circRNAs that modulate NDV replication through autophagy and elucidate their underlying mechanisms. Our findings revealed that circRNA0857 was significantly upregulated in NDV-infected cells. Functional assays showed that silencing circRNA0857 markedly inhibited NDV replication, while its overexpression enhanced viral replication. Mechanistic analyses demonstrated that circRNA0857 functions as a “miRNA sponge,” specifically targeting and sequestering miR1709 and miR1746. These miRNAs were found to regulate the expression of ATG3 and ATG7, respectively, two key autophagy-related genes in the autophagy pathway. By sponging miR1709 and miR1746, circRNA0857 upregulates the expression of ATG3 and ATG7, thereby enhancing autophagy and promoting NDV infection. This study is the first to identify circRNA0857 as a critical regulator in NDV infection, elucidating its role in enhancing NDV-induced autophagy via the miR1709/ATG3 and miR1746/ATG7 signaling axes. These findings significantly advance our understanding of how circRNAs regulate viral infections and provide novel insights into the molecular mechanisms underlying NDV replication. IMPORTANCE Circular RNAs (circRNAs), as endogenous non-coding RNAs, are widely involved in various biological processes, particularly in the regulatory modulation of miRNA functions. Newcastle disease virus (NDV) can replicate in a variety of cells and promote its proliferation by hijacking and utilizing autophagy. In this study, we screened for novel circular RNAs that regulate NDV replication through modulating host autophagy and identified circRNA0857 as a key regulator of NDV infection. It enhances autophagy and promotes viral replication by acting as a “miRNA sponge,” simultaneously targeting the two signaling axes miR1709/ATG3 and miR1746/ATG7. This research fills a gap in understanding how circRNAs regulate NDV-induced autophagy and provides a new perspective for understanding the interactions between host non-coding RNAs and viruses. This finding deepens our understanding of the pathogenic mechanisms of NDV and offers potential molecular targets for the development of antiviral strategies based on circular RNAs or miRNAs. Circular RNAs (circRNAs), as endogenous non-coding RNAs, are widely involved in various biological processes, particularly in the regulatory modulation of miRNA functions. Newcastle disease virus (NDV) can replicate in a variety of cells and promote its proliferation by hijacking and utilizing autophagy. In this study, we screened for novel circular RNAs that regulate NDV replication through modulating host autophagy and identified circRNA0857 as a key regulator of NDV infection. It enhances autophagy and promotes viral replication by acting as a “miRNA sponge,” simultaneously targeting the two signaling axes miR1709/ATG3 and miR1746/ATG7. This research fills a gap in understanding how circRNAs regulate NDV-induced autophagy and provides a new perspective for understanding the interactions between host non-coding RNAs and viruses. This finding deepens our understanding of the pathogenic mechanisms of NDV and offers potential molecular targets for the development of antiviral strategies based on circular RNAs or miRNAs.
Siyuan Wang, Yang Qu, Weiwei Liu et al.· Journal of Virology· 0 citations
Newcastle disease virus (NDV), a significant avian paramyxovirus, depends on the acquisition of host-derived membranes for viral envelope assembly during budding. However, the relationship between NDV budding efficiency and host metabolic reprogramming remains incompletely understood. Our previous research demonstrated that the highly virulent strain Herts/33 and the non-virulent strain LaSota differ significantly in budding efficiency, which is attributed to the difference in ubiquitination levels at the K247 site of the M protein. In this study, using recombinant viruses generated in our previous study, we investigated the effects of the K247 ubiquitination site on host cell metabolism. Through RNA sequencing and LC–MS/MS, systematically profiled the transcriptional and metabolic alterations induced by these viruses in HeLa cells. We also compared organ pathology in three-week-old SPF chicks infected with Herts/33 versus LaSota. The results showed that NDV infection broadly reprograms host sphingolipid metabolism. Compared with rLaSota-WT, rLaSota-R247K exhibited higher budding efficiency, linked to accelerated ceramide depletion and a stronger innate immune response. Specifically, rLaSota-R247K infection upregulated interferon- and interleukin-related components such as ISG15, CXCL8, TNF-α, CXCL10, NOD2, CD274, OAS, and IFNB1. Furthermore, we confirmed that blocking the ceramide synthesis pathway significantly suppresses NDV-M protein-mediated budding of VLPs and virions. Pathologically, Herts/33 induced more severe tissue damage than LaSota. Together, these findings indicate that the K247 residue of the NDV M protein enhances viral budding and promotes rapid utilization of host sphingolipids. This work provides mechanistic insight into NDV budding and highlights virus–host metabolic interactions that may inform future antiviral strategies.
Jun Dai, Fan-Xin Liu, Yiyi Feng et al.· Poultry Science· 0 citations
The Importin α family, as key mediators of nucleocytoplasmic transport, represents a common target for viral immune evasion. However, whether coronaviruses directly manipulate Importin α to disrupt nuclear trafficking and suppress antiviral immunity has remained unclear. In this study, we identify a previously unrecognized mechanism by which coronaviruses from all four genera subvert host innate immunity through the proteolytic inactivation of Importin α1, a key mediator of nuclear import of cargo proteins. We demonstrate that the membrane-associated papain-like protease (PLpro-TM) directly cleaves Importin α1 at specific glycine residues, G129 for PEDV and IBV PLpro, and G119 for MHV and PDCoV PLpro, thereby disrupting its nuclear import function. This cleavage impairs the nuclear translocation of multiple transcription factors (IRF3, STAT1, STAT2, and p65) and suppresses the expression of downstream antiviral genes, including IFN-β and IFN-stimulated genes (ISGs). Importantly, cleavage-resistant mutants of Importin α1 (G129A or G119A) restore nuclear import capability and rescue IFN-β signaling. Consequently, they exert a more potent inhibitory effect on viral replication than the wild-type Importin α1, fulfilling an antiviral role. Our work establishes PLpro-TM-mediated cleavage of Importin α1 as a conserved immune evasion strategy across coronaviruses and highlights this interaction as a potential target for broad-spectrum antiviral intervention. Author summary The nuclear transport of transcription factors is a critical checkpoint for the initiation of innate antiviral immunity. Here, we identify the PLpro-TM protein as a pan-coronavirus antagonist of nucleocytoplasmic trafficking and innate immune response. We demonstrate that PLpro-TM from four distinct coronavirus genera directly cleaves Importin α1 at specific glycine residues, thereby disabling its ability to mediate the nuclear import of key transcription factors and subsequent transcription of anti-viral genes. This work reveals a previously unrecognized, evolutionarily conserved immune evasion strategy shared across α, β, γ, and δ coronaviruses. By uncovering the proteolytic inactivation of Importin α1 targeting by PLpro-TM, our findings not only resolve a long-standing question about how coronaviruses disrupt nuclear trafficking, but also establish PLpro-TM and Importin α1 as promising targets for the development of broad-spectrum antiviral therapeutics against current and emerging coronaviruses.
Jiehuang Wang, Wenxiang Xue, Yingjie Sun et al.· bioRxiv· 0 citations
We use cookies to run the site and, with your consent, for analytics and to show ads.
See our Cookie Policy.
