title: Molecular cloning and functional characterization of porcine cyclic GMP-AMP synthase.
abstract: Cyclic GMP-AMP synthase (cGAS), which belongs to the nucleotidyltransferase family, recognizes cytosolic DNA and induces the type I interferon (IFN) pathway through the synthesis of the second messenger cGAMP. In this study, porcine cGAS (p-cGAS) was identified and its tissue distribution, subcellular localization, and functions in innate immunity were characterized. The coding sequence of p-cGAS is 1494 bp long, encodes 497 amino acids, and is most similar (74%) to Bos taurus cGAS. p-cGAS mRNA is abundant in the spleen, duodenum, jejunum, and ileum. The subcellular distribution of p-cGAS is not only in the cytosol, but also on the endoplasmic reticulum (ER) membrane. The overexpression of wild-type p-cGAS in porcine kidney epithelial cells, but not its catalytically inactive mutants, induced IFN-β expression, which was dependent on STING and IRF3. However, the downregulation of p-cGAS by RNA interference markedly reduced IFN-β expression after pseudorabies virus (PRV) infection or poly(dA:dT) transfection. These results demonstrate that p-cGAS is an important DNA sensor, required for IFN-β activation.
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China; Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Wang
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China; Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China. Electronic address: chubeibei_hau@hotmail.com.
authors.forename: Beibei
authors.initials: B
authors.lastname: Chu
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China; Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China.
authors.forename: Lili
authors.initials: L
authors.lastname: Du
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China; Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China.
authors.forename: Yingqian
authors.initials: Y
authors.lastname: Han
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China; Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China; Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China.
authors.forename: Shuangshuang
authors.initials: S
authors.lastname: Fan
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China; Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China.
authors.forename: Yueying
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China; Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou 450002, Henan Province, PR China. Electronic address: haubiochem@163.com.
authors.forename: Guoyu
authors.initials: G
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003001:Cloning, Molecular; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000276:immunology; D005786:Gene Expression Regulation / Q000276:immunology*; D016899:Interferon-beta / Q000235:genetics / Q000276:immunology; D007425:Intracellular Membranes / Q000276:immunology*; D008969:Molecular Sequence Data; D009154:Mutation; D009713:Nucleotidyltransferases / Q000235:genetics / Q000276:immunology; D009928:Organ Specificity / Q000235:genetics / Q000276:immunology; D011067:Poly dA-dT / Q000494:pharmacology; D017386:Sequence Homology, Amino Acid; D013552:Swine / Q000235:genetics / Q000276:immunology
chemical_list: D011067:Poly dA-dT; D016899:Interferon-beta; D009713:Nucleotidyltransferases
keywords: Interferon; PRV; Porcine; cGAS