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Functional characterization of porcine LSm14A in IFN-β induction
Guilan Wen1, Yi Zhang, Xian Zhang
1Zhejiang University Institute of Preventive Veterinary Medicine, Zhejiang Province Key Laboratory of Preventive Veterinary Medicine, Hangzhou, China.
Abstract:
Human LSm14A has recently been found as a processing body-associated sensor of intracellular viral nucleic acids and triggers signaling for type I IFN expression. Here porcine LSm14A (pLSm14A) was cloned from the PK-15 cells. The pLSm14A ORF is 1392 bp in length, encoding 463 amino acids. The putative pLSm14A contains a Sm-like domain and two arginine-glycine-glycine (RGG) boxes. The pLSm14A has high identity at the amino acid level to those of bovine, human and mouse (93.5-97.4%) and is transcribed in different tissues of pigs. In HEK293 or Marc-145 cells, pLSm14A was localized in the cytosol as P-body-like dots. Expression of pLSm14A in HEK293 or Marc-145 cells enhanced activities of IFN-β and NF-κB promoters, induced IFN-β transcription, and potentiated poly(I:C)-induced IFN-β promoter activation, indicating that pLSm14A is a potential signal molecule in the IFN-β pathway of pigs. We also found that pLSm14A-induced IFN-β promoter activity was down-regulated by porcine reproductive and respiratory syndrome virus infection in Marc-145 cells. Since pLSm14A is constitutively expressed in virtually all tissues, more research is needed to explore its role in initial phase of viral infections of pigs and its relationship with RIG-I in sensing PAMPs for type I IFN induction.
Insights
Porcine LSm14A acts as a sensor for viral nucleic acids, promoting type I interferon (IFN) expression. Its activity is reduced by porcine reproductive and respiratory syndrome virus, highlighting its role in antiviral defense.
Area of Science:
- Immunology
- Virology
- Molecular Biology
Background:
- Human LSm14A functions as a sensor for intracellular viral nucleic acids, initiating type I interferon (IFN) signaling.
- Understanding the role of LSm14A in pigs is crucial for swine antiviral immunity.
Purpose of the Study:
- To clone and characterize porcine LSm14A (pLSm14A).
- To investigate the function of pLSm14A in the type I IFN pathway in pigs.
- To explore the impact of porcine reproductive and respiratory syndrome virus (PRRSV) on pLSm14A activity.
Main Methods:
- Cloning of pLSm14A from PK-15 cells.
- Sequence analysis and identification of functional domains (Sm-like, RGG boxes).
- Subcellular localization studies in HEK293 and Marc-145 cells.
- Reporter gene assays to measure IFN-β and NF-κB promoter activity.
- Assessment of pLSm14A response to poly(I:C) stimulation.
- Evaluation of PRRSV infection effects on pLSm14A-induced promoter activity.
Main Results:
- Porcine LSm14A (pLSm14A) was cloned, encoding 463 amino acids with conserved domains.
- pLSm14A localized to cytosol as P-body-like dots in mammalian cells.
- pLSm14A enhanced IFN-β and NF-κB promoter activities and IFN-β transcription.
- pLSm14A potentiated poly(I:C)-induced IFN-β promoter activation.
- PRRSV infection down-regulated pLSm14A-induced IFN-β promoter activity.
Conclusions:
- pLSm14A functions as a potential signal molecule in the pig's type I IFN pathway.
- pLSm14A plays a role in sensing viral nucleic acids and inducing IFN-β.
- PRRSV infection interferes with pLSm14A's antiviral signaling.
- Further research is needed on pLSm14A's role in early viral infections and its interaction with RIG-I for type I IFN induction.
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