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Updated: Apr 14, 2026

MicroRNA-based Regulation of Picornavirus Tropism
Published on: February 6, 2017
Pestiviruses utilize pyrimidine metabolism to regulate mitophagy for viral replication
Bingqian Zhao1,2,3, Jing Chen1, Yan Cheng1,2
1MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.
None:
DHODH (dihydroorotate dehydrogenase (quinone)) has been demonstrated as a critical regulator of programmed cell death, yet its role in macroautophagy/autophagy remains poorly defined. Flaviviridae pose a significant threat to global public health, and their replication is closely associated with autophagy. Building upon our previous findings that DHODH was a broad-spectrum target for Flaviviridae and a key regulator of Pestiviruses replication, this study employed RNA-seq screening coupled with functional validation to demonstrate that DHODH affected Pestiviruses replication by regulating mitophagy. Notably, we observed remarkable virus genus specificity in this regulatory mechanism. For autophagy-dependent Pestiviruses, DHODH deficiency impaired autophagosome-lysosome fusion, thereby suppressing viral replication. Conversely, in autophagy-inhibiting Flaviviruses, the blockade of autophagy flux facilitated viral replication. These observations underscore the specificity of DHODH-mediated viral replication regulation. Additionally, compound supplementation assays indicated that DHODH regulated autophagy via pyrimidine nucleotide metabolism, as exogenous pyrimidine precursors restored autophagosome-lysosome fusion. Furthermore, our research uncovered a novel mechanism whereby classical swine fever virus (CSFV) non-structural protein 4A (NS4A) recruited DHODH to mitochondria, facilitating its interaction with MAP1LC3/LC3 (microtubule associated protein 1 light chain 3) through the LC3-interacting region (LIR) domain to activate mitophagy. Collectively, our findings highlight DHODH as a promising antiviral target within the metabolism-autophagy axis, providing novel insights for antiviral drug development.Abbreviation: AMPK: AMP-activated protein kinase; ATF4: activating transcription factor 4; ATG5: autophagy related 5; BafA1: bafilomycin A1; BNIP3L/NIX: BCL2 interacting protein 3 like; BVDV: bovine viral diarrhea virus; CALCOCO2/NDP52: calcium binding and coiled-coil domain 2; CCCP: carbonyl cyanide 3-chlorophenylhydrazone; co-IP: co-immunoprecipitation; COX4: cytochrome c oxidase subunit 4; CQ: chloroquine; CSFV: classical swine fever virus; DAPI: 4',6-diamidino-2-phenylindole; DEGs: differentially expressed genes; DHO: DHODH substrate dihydroorotate; DHODH: dihydroorotate dehydrogenase; DTMUV: duck tembusu virus; FIS1: fission mitochondrial 1; FUNDC1: FUN14 domain containing 1; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GFP: green fluorescent protein; GO: gene ontology; HSPA/HSP70: heat shock protein family A (Hsp70); JEV: Japanese encephalitis virus; KEGG: kyoto encyclopedia of genes and genomes; LIR: LC3-interacting region; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; Mdivi-1: mitochondrial division inhibitor 1; MFF: mitochondrial fission factor; MFN1: mitofusin 1; MFN2: mitofusin 2; MITO: mitochondria; MOI: multiplicity of infection; MTOR: mechanistic target of rapamycin kinase; MTS: mitochondrial targeting signal; OPTN: optineurin; ORO: DHODH product orotate; PBS: phosphate-buffered saline; PRKN: parkin RBR E3 ubiquitin protein ligase; PYR: pyrazofurin; RAPA: rapamycin; RFP: red fluorescent protein; RNA-seq: RNA sequencing; RT-qPCR: reverse transcription-quantitative real-time polymerase chain reaction; SD: standard deviation; siRNA: small interfering RNA; SQSTM1/p62: sequestosome 1; TOMM20: translocase of outer mitochondrial membrane 20; UMP: uridine monophosphate; VDAC1: voltage dependent anion channel 1.
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