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Cis-acting structural element in 5' UTR is essential for infectivity of porcine reproductive and respiratory syndrome virus.

ABSTRACT: It is believed that the genomic 5' untranslated region (UTR) of Arterivirus plays crucial roles in viral genomic replication, subgenomic mRNA transcription and protein translation, yet the structure and function still remain largely unknown. In this study, we conducted serial nucleotide truncation, ranging from 1 to 190 nucleotides, to the 5' UTR of the porcine reproductive and respiratory syndrome virus (PRRSV) infectious full-length cDNA clone pAPRRS. In vitro synthetic RNAs were transfected into MARC-145 cells for further genetic and virologic analysis. Our results demonstrated that the first three nucleotides of PRRSV 5' UTR were dispensable for virus viability, which however was repaired with foreign sequences. In order to assess if the primary sequence or structural element play more important regulatory roles, the CMV promoter-driven 5' UTR truncation mutant cDNA clones were directly transfected into the BHK-21 cell lines. We found that PRRSV tolerated the first 16 nucleotides sequence alteration of 5' UTR without losing virus viability. However, these revertant viruses contained a range of non-templated with unknown origin exogenous nucleotides in the repaired 5' end. Further analyses revealed that the 5' proximal stem-loop 1 (SL1) in the highly structured 5' UTR was invariably required for virus infectivity. Taken together, we conclude that authentic 5'-proximal primary sequence is nonessential, but the resultant structural elements are probably indispensable for PRRSV infectivity.

SUBMITTER: Gao F

PROVIDER: S-EPMC7114472 | biostudies-literature | 2012 Jan

REPOSITORIES: biostudies-literature

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Cis-acting structural element in 5' UTR is essential for infectivity of porcine reproductive and respiratory syndrome virus.

Gao Fei F Lu Jiaqi J Yao Huochun H Wei Zuzhang Z Yang Qian Q Yuan Shishan S

Virus research 20110908 1

It is believed that the genomic 5' untranslated region (UTR) of Arterivirus plays crucial roles in viral genomic replication, subgenomic mRNA transcription and protein translation, yet the structure and function still remain largely unknown. In this study, we conducted serial nucleotide truncation, ranging from 1 to 190 nucleotides, to the 5' UTR of the porcine reproductive and respiratory syndrome virus (PRRSV) infectious full-length cDNA clone pAPRRS. In vitro synthetic RNAs were transfected i ...[more]

PMID: 21924304

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Project description:BackgroundThe porcine reproductive and respiratory syndrome (PRRS) is a devastating disease for the pig industry. In this study, we analysed the genetic variability of PRRS virus (PRRSV) as well as the relationship between the genetic variability, the geographical and temporal distribution of the PRRSV strains. Moreover, we investigated the association between the glycosylation patterns in PRRSV sequences and pigs growth.ResultsThe data highlight that PRRSV strains evolve rapidly on individual farms, and temporal evolution of PRRSV is an important factor of genetic variability. Analysis of glycosylation sites in the glycoprotein 5 (GP5) ectodomain revealed that PRRSV isolates had seven combinations of putative N-linked glycosylation sites of which the N37/46/53 sites was found in 79% of the sequences. No significant relationship was found between the genetic variation of the PRRSV strains and the geographic distance. A significant relationship was found between the genetic variation and time of sampling when farm was considered as a factor in the analysis. Furthermore, the commercial semen from artificial insemination centres was not a source of PRRS transmission.The PRRSV having the glycosylation site at position N46 (N46+) were observed to have higher burden on pigs and accordingly the corresponding infected pigs had lower average daily gain (ADG) compared with those infected with PRRSV lacking the glycosylation at N46 (N46-) position site. This study showed that the number of piglets by litter infected by PRRSV was lower for the Landrace breed than for the other studied breeds (Large White, Duroc and Pietrain).ConclusionsThe PRRSV genetic variability which is determined by a local and temporal evolution at the farm level could be considered in a perspective of prevention. Moreover, the association between the PRRSV glycosylation patterns and its virulence could be of interest for vaccine development. The differences of resistance to PRRSV infections among pig breeds might open new horizons for the genetic selection of robustness against PRRSV infection.

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Cis-acting structural element in 5' UTR is essential for infectivity of porcine reproductive and respiratory syndrome virus.

Publications

Cis-acting structural element in 5' UTR is essential for infectivity of porcine reproductive and respiratory syndrome virus.

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