Hyper-phosphorylation of nsp2-related proteins of porcine reproductive and respiratory syndrome virus

Shang, Peng; Yuan, Fuh-Gwo; Misra, Saurav; Lǐ, Yànhuá; Fāng, Yīng

doi:10.1016/j.virol.2020.01.018

Cited by 4 publications

(9 citation statements)

References 73 publications

(155 reference statements)

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“…In PRRSV, it has been proven that aspartic 185 of the Nsp4 could regulate the capacity to antagonize IFN-I expression and lead to a slower replication rate for PRRSV [ 14 ]; Mutations to serine 519 , threonine 544 , threonine 586 and serine 592 of Nsp9 have proved to affect the replication ability and virulence of high pathogenicity (HP) –PRRSV [ 16 ]. Serine 918 of the Nsp2 has proved to regulate virus production [ 26 ].…”

Section: Discussionmentioning

confidence: 99%

A novel amino acid site of N protein could affect the PRRSV-2 replication by regulating the viral RNA transcription

et al. 2022

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Background Finding the key amino acid sites that could affect viral biological properties or protein functions has always been a topic of substantial interest in virology. The nucleocapsid (N) protein is one of the principal proteins of the porcine reproductive and respiratory syndrome virus (PRRSV) and plays a vital role in the virus life cycle. The N protein has only 123 or 128 amino acids, some of key amino acid sites which could affect the protein functions or impair the viral biological characteristics have been identified. In this research, our objective was to find out whether there are other novel amino acid sites of the N protein can affect N protein functions or PRRSV-2 replication. Results In this study, we found mutated the serine78 and serine 99of the nucleocapsid (N) protein can reduce the N-induced expression of IL-10 mRNA; Then, by using reverse genetics system, we constructed and rescued the mutant viruses, namely, A78 and A99.The IFA result proved that the mutations did not affect the rescue of the PRRSV-2. However, the results of the multistep growth kinetics and qPCR assays indicated that, compared with the viral replication ability, the titres and gRNA levels of A78 were significantly decreased compared with the wild-type. Further study showed that a single amino acid change from serine to alanine at position 78 of the N protein could abrogates the level of viral genomic and subgenomic RNAs. It means the mutation could significant decrease the viral replication efficiency in vitro. Conclusions Our results suggest that the serine78 of N protein is a key site which could affect the N protein function and PRRSV replication ability.

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Section: Discussionmentioning

confidence: 99%

A novel amino acid site of N protein could affect the PRRSV-2 replication by regulating the viral RNA transcription

et al. 2022

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“…Two PCR reactions (PCR1 and PCR2) were performed to amplify the genomic region between XbaI and NotI. The overlapping part of the 3′ end of the To precisely quantify the ratios of gRNA and sgRNA (plus strand or minus strand), RT-qPCR was conducted with a panel of specific primers/probe set, which were designed to cover the leader-B-TRS-junction sites as we described previously (30). For a nucleotide sequence in the TRS of sgRNA2, 6, and 7 that shown heterogeneity, a degenerate nucleotide was used in the probes (30).…”

Section: Construction and Production Of Prrsv Mutantsmentioning

confidence: 99%

“…; https://doi.org/10.1101/2024.05. 30.596123 doi: bioRxiv preprint antisense (−) B-TRS with a sense (+) L-TRS. Every B-TRS would serve as an attenuation signal to polymerase processivity, with (−) RNA synthesis either able to continue on the (+) gRNA template, or else the nascent (−) RNA disassociates from the (+) gRNA template at the B-TRS and reanneals to a (+) gRNA template at the L-TRS, where polymerase activity resumes to synthesize the antisense leader sequence templated by the 5′-proximal region of the (+) gRNA.…”

Section: Introductionmentioning

confidence: 99%

“…; https://doi.org/10.1101/2024.05. 30.596123 doi: bioRxiv preprint polyprotein processing remain unaltered (15,16). Additionally, mutagenesis studies revealed that the EAV nsp1 protein is a prominent trans-acting factor to ensure the equilibrium of sgRNA transcription and genome replication, and maintaining the production ratios of different sgRNA species (14).…”

Section: Introductionmentioning

confidence: 99%

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An intra-family conserved high-order RNA structure within the M ORF is important for arterivirus subgenomic RNA accumulation and infectious virus production

Shang,

Li,

Chen

et al. 2024

Preprint

Self Cite

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Synthesis of subgenomic RNAs is a strategy commonly used by polycistronic positive sense single-stranded RNA viruses to express 3′-proximal genes. Members of the order ofNidovirales, including coronaviruses and arteriviruses, use a unique discontinuous transcription strategy to synthesize subgenomic RNAs. In this study,in silicosynonymous site conservation analysis and RNA structure folding predicted the existence of intra-family conserved high-order RNA structure within the M ORF of arteriviral genomes, which was further determined to be important for the transcription/accumulation of subgenomic RNAs and production of infectious viral particles. Mutations disrupting the stability of the RNA structures significantly decreased the accumulation of multiple subgenomic RNAs. In contrast, the impact of mutagenesis on full-length genomic RNA accumulation was limited. The degree to which wild-type levels of subgenomic RNA accumulation were maintained was found to correlate with the efficiency of infectious virus production. Moreover, the thermo-stability of stems within the high-order RNA structure is also well correlated with viral replication capacity and the maintenance of subgenomic RNA accumulation. This study is the first to report an intra-Arteriviridaeconserved high-order RNA structure that is located in a protein-coding region and functions as an importantcis-acting element to control the accumulation/transcription of arteriviral subgenomic RNAs. This work suggests a complex regulation mechanism between genome replication and discontinuous transcription in nidoviruses.IMPORTANCEArteriviruses are a group of RNA viruses that infect different animal species. They can cause diseases associated with respiratory/reproductive syndromes, abortion, or haemorrhagic fever. Among arteriviruses, porcine reproductive and respiratory syndrome virus (PRRSV) and equine arteritis virus (EAV) are economically important veterinary pathogens. The challenge in control of arterivirus infection reflects our limited knowledge of viral biology. In this study, we conducted a comprehensive bioinformatical analysis of arteriviral genomes and discovered intra-family conserved regions in the M ORF with a high-order RNA structure. The thermo-stability of the RNA structure influences sgRNA transcription/accumulation and correlates with the level of infectious virus production. Our studies provide a new insight on arterivirus replication mechanism, which may have implications in developing disease control and prevention strategies.

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“…The 3' terminal ORFs2-7 code for structural proteins (e.g., GP2a, E, GP3, GP4, GP5, M and N, etc.) [6,7], whereas ORF1a and ORF1b account for nearly 75% of the 5'-end of the genome and encode viral two important viral replicase polyproteins (pp1a and pp1ab) that are further processed into at least 16 nonstructural replicase proteins (nsps). These nsps then come together to form viral replication and transcription complex (RTC) for genome replication and subgenomic RNA synthesis.…”

Section: Introductionmentioning

confidence: 99%

PRRSV Non-Structural Proteins Orchestrate Porcine E3 Ubiquitin Ligase RNF122 to Promote PRRSV Proliferation

Sun

Guo

et al. 2022

Viruses

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Ubiquitination plays a major role in immune regulation after viral infection. An alternatively spliced porcine E3 ubiquitin ligase RNF122 promoted PRRSV infection and upregulated in PRRSV-infected PAM cells was identified. We characterized the core promoter of RNF122, located between −550 to −470 bp upstream of the transcription start site (TSS), which displayed significant differential transcriptional activities in regulating the transcription and expression of RNF122. The transcription factor HLTF was inhibited by nsp1α and nsp7 of PRRSV, and the transcription factor E2F complex regulated by nsp9. Together, they modulated the transcription and expression of RNF122. RNF122 could mediate K63-linked ubiquitination to raise stability of PRRSV nsp4 protein and thus promote virus replication. Moreover, RNF122 also performed K27-linked and K48-linked ubiquitination of MDA5 to degrade MDA5 and inhibit IFN production, ultimately promoted virus proliferation. In this study, we illustrate a new immune escape mechanism of PRRSV that enhances self-stability and function of viral nsp4, thus, regulating RNF122 expression to antagonize IFNα/β production. The present study broadens our knowledge of PRRSV-coding protein modulating transcription, expression and modification of host protein to counteract innate immune signaling, and may provide novel insights for the development of antiviral drugs.

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Hyper-phosphorylation of nsp2-related proteins of porcine reproductive and respiratory syndrome virus

Cited by 4 publications

References 73 publications

A novel amino acid site of N protein could affect the PRRSV-2 replication by regulating the viral RNA transcription

A novel amino acid site of N protein could affect the PRRSV-2 replication by regulating the viral RNA transcription

An intra-family conserved high-order RNA structure within the M ORF is important for arterivirus subgenomic RNA accumulation and infectious virus production

PRRSV Non-Structural Proteins Orchestrate Porcine E3 Ubiquitin Ligase RNF122 to Promote PRRSV Proliferation

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