Molecular cloning and functional analysis of Macaca mulatta STING

Area of Science:

• Immunology
• Molecular Biology
• Primate Genetics

Background:

• Stimulator of Interferon Genes (STING) is a key immune adaptor molecule involved in various immune responses.
• STING has been characterized in several species, but not yet in non-human primates.
• Understanding STING in primates is crucial for comparative immunology and disease research.

Purpose of the Study:

• To clone and characterize the Stimulator of Interferon Genes (STING) gene in rhesus macaques (Macaca mulata).
• To investigate the functional role of M. mulatta STING in the interferon (IFN) pathway.
• To establish a foundation for understanding primate immune responses.

Main Methods:

• Cloning of the M. mulatta STING complementary DNA.
• Phylogenetic analysis comparing M. mulatta STING with other species.
• Functional assays measuring promoter activities (IFN-β, NF-κB, ISRE) and mRNA levels (IFN-α, IFN-β, IRF3).
• Cell-based assays using Marc-145 cells infected with PRRSV or treated with poly (I:C) and poly (dA:dT).

Main Results:

• The M. mulatta STING cDNA is 1140 bp, encoding 380 amino acids.
• Phylogenetic analysis indicates a close evolutionary relationship between human and rhesus macaque STING.
• M. mulatta STING significantly enhanced IFN-β, NF-κB, and ISRE promoter activities.
• STING overexpression increased mRNA levels of IFN-α, IFN-β, and IRF3.
• Porcine reproductive and respiratory syndrome virus infection activated STING in Marc-145 cells in a time- and multiplicity-dependent manner.
• Poly (I:C) and poly (dA:dT) treatments upregulated STING expression in a time- and dose-dependent manner.

Conclusions:

• Rhesus macaque STING is a vital signaling protein in the interferon pathway.
• This study provides the first characterization of STING in non-human primates.
• The findings contribute to understanding rhesus macaque immunity and have implications for primate and human disease research.