Grass carp (Ctenopharyngodon idella) GPATCH3 initiates IFN 1 expression via the activation of STING-IRF7 signal axis

Li, Meifeng; Shi, Yang; Xu, Xiaowen; Liu, Yapeng; Jiang, Zeying; Li, Yinping; Lv, Yangfeng; Lu, Shina; Hu, Chengyu; Mao, Huiling

doi:10.1016/j.dci.2020.103781

Cited by 9 publications

(8 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Due to the high conservation between grass carp and human KAT8 (86%), anti-human KAT8 rabbit monoclonal antibody (HUABIO, China) was used to detect grass carp KAT8. Anti-IRF7 Ab (1:1000) was from HUABIO Biological and used as previously described (37). GFP-Tag and Flag-Tag Ab (1:5000) were purchased from Sigma (USA) and Abmart (USA), respectively.…”

Section: Methodsmentioning

confidence: 99%

“…The ORF of CiKAT8, CiIRF3 and CiIRF7 were separately inserted into pcDNA3.1 (+) vector (Invitrogen, USA) for over-expression experiments. IFN 1 and ISG15 promoter luciferase reporter plasmids were prepared in our lab (37); pISRE-TA-luc (beyotime, China) was used in luciferase assays. Each construction was confirmed by DNA sequencing.…”

Section: Methodsmentioning

confidence: 99%

“…CIK cells transfected with 2 µg oligomeric nucleic acids [poly(I: C) or B-DNA or Z-DNA] or 50 µl of 10 −8 TCID 50 viruses (GCRV or SVCV) were separately cultured in 6-well plates for 6, 12, 24, 48, and 72 h, as described in our previous studies (37)(38)(39).…”

Section: The Expression Profile Of Kat8 and Quantitative Real-time Pcr (Qrt-pcr)mentioning

confidence: 99%

See 2 more Smart Citations

Grass Carp (Ctenopharyngodon idella) KAT8 Inhibits IFN 1 Response Through Acetylating IRF3/IRF7

Mao

et al. 2022

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

Post-translational modifications (PTMs), such as phosphorylation and ubiquitination, etc., have been reported to modulate the activities of IRF3 and IRF7. In this study, we found an acetyltransferase KAT8 in grass carp (CiKAT8, MW286472) that acetylated IRF3/IRF7 and then resulted in inhibition of IFN 1 response. CiKAT8 expression was up-regulated in the cells under poly I:C, B-DNA or Z-DNA stimulation as well as GCRV(strain 873) or SVCV infection. The acetyltransferase domain (MYST domain) of KAT8 promoted the acetylation of IRF3 and IRF7 through the direct interaction with them. So, the domain is essential for KAT8 function. Expectedly, KAT8 without MYST domain (KAT8-△264-487) was granularly aggregated in the nucleus and failed to down-regulate IFN 1 expression. Subcellular localization analysis showed that KAT8 protein was evenly distributed in the nucleus. In addition, we found that KAT8 inhibited the recruitment of IRF3 and IRF7 to ISRE response element. Taken together, our findings revealed that grass carp KAT8 blocked the activities of IRF3 and IRF7 by acetylating them, resulting in a low affinity interaction of ISRE response element with IRF3 and IRF7, and then inhibiting nucleic acids-induced innate immune response.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Grass Carp (Ctenopharyngodon idella) KAT8 Inhibits IFN 1 Response Through Acetylating IRF3/IRF7

Mao

et al. 2022

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Finally, the membranes were exposed using a chemiluminescence imaging system (CLINX, China). Specific antibodies used here are against grass carp type I IFN (1:800, saved in our lab) ( 28 ), phosphorylated IRF3 (p-IRF3; 1:1,000, Beyotime Biotechnology, China, AF1594) ( 27 ), basal and phosphorylated IRF7 (p-IRF7; 1:1,000, HuaBio, China, ET1610-89) ( 29 ), eIF2α (1:500, saved in our lab) ( 30 ), phosphorylated eIF2α (p-eIF2α; 1:1,000, Abcam, UK, ab32157) ( 30 ), and GAPDH (loading control; 1:10,000, saved in our lab) ( 30 ). All commercial antibodies against p-IRF3, IRF7/p-IRF7, and p-eIF2α cross-react with grass carp, which is confirmed by transfection of pEGFP-IRF3, -IRF7, and -eIF2α into grass carp cells followed by comparison of the size of the basal and GFP recombinant proteins of each gene in our previous work ( 27 , 29 , 30 ).…”

Section: Methodsmentioning

confidence: 99%

Fish Paralog Proteins RNASEK-a and -b Enhance Type I Interferon Secretion and Promote Apoptosis

Sun

Jiang

et al. 2021

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

Type I interferon and apoptosis elicit multifaceted effects on host defense and various diseases, such as viral infections and cancers. However, the gene/protein network regulating type I interferon and apoptosis has not been elucidated completely. In this study, we selected grass carp (Ctenopharyngodon idella) as an experimental model to investigate the modulation of RNASEK on the secretion of type I interferon and apoptosis. We first cloned two paralogs RNASEK-a and -b in grass carp, defined three exons in each gene, and found the length of both coding regions is 306 bp with 73.27% of protein homology. The protein sequences of the two paralogs are highly conserved across species. Two proteins were mainly localized in early and late endosomes and endoplasmic reticulum. Further, quantitative real-time PCR demonstrated that dsRNA poly I:C and grass carp reovirus upregulated RNASEK-a and -b in grass carp cells and tissues. Overexpression of RNASEK-a and -b individually induced type I interferon expression and the phosphorylation of IRF3/IRF7 shown by Western blot and immunofluorescent staining, increased Bax/Bcl-2 mRNA ratio, DNA fragmentations, TUNEL-positive cells, and the proportion of Annexin V-positive signals in flow cytometry, and activated eIF2α, opposite to that observed when RNASEK-a and -b were knocked down in multiple cell types. Taken together, we claim for the first time that fish paralog proteins RNASEK-a and -b enhance type I interferon secretion and promote apoptosis, which may be involved in the phosphorylation of IRF3/IRF7 and eIF2α, respectively. Our study reveals a previously unrecognized role of RNASEK as a new positive regulator of type I interferon and apoptosis.

show abstract

“…Specific antibodies used here are against grass carp type I IFN (1:800, saved in our lab) (28), phosphorylated IRF3 (p-IRF3; 1:1,000, Beyotime Biotechnology, China, AF1594) (27), basal and phosphorylated IRF7 (p-IRF7; 1:1,000, HuaBio, China, ET1610-89) (29), eIF2a (1:500, saved in our lab) (30), phosphorylated eIF2a (p-eIF2a; 1:1,000, Abcam, UK, ab32157) (30), and GAPDH (loading control; 1:10,000, saved in our lab) (30). All commercial antibodies against p-IRF3, IRF7/p-IRF7, and p-eIF2a cross-react with grass carp, which is confirmed by transfection of pEGFP-IRF3, -IRF7, and -eIF2a into grass carp cells followed by comparison of the size of the basal and GFP recombinant proteins of each gene in our previous work (27,29,30).…”

Section: Western Blotmentioning

confidence: 70%

DataSheet_1.docx

View full text Add to dashboard Cite

Grass carp (Ctenopharyngodon idella) GPATCH3 initiates IFN 1 expression via the activation of STING-IRF7 signal axis

Cited by 9 publications

References 36 publications

Grass Carp (Ctenopharyngodon idella) KAT8 Inhibits IFN 1 Response Through Acetylating IRF3/IRF7

Grass Carp (Ctenopharyngodon idella) KAT8 Inhibits IFN 1 Response Through Acetylating IRF3/IRF7

Fish Paralog Proteins RNASEK-a and -b Enhance Type I Interferon Secretion and Promote Apoptosis

DataSheet_1.docx

Contact Info

Product

Resources

About