Abstract
Autophagy is regulated for IFN-gamma-mediated antimicrobial efficacy; however, its molecular effects for IFN-gamma signaling are largely unknown. Here, we show that autophagy facilitates IFN-gamma-activated Jak2-STAT1. IFN-gamma induces autophagy in wild-type but not in autophagy protein 5 (Atg5(-/-))-deficient mouse embryonic fibroblasts (MEFs), and, autophagy-dependently, IFN-gamma induces IFN regulatory factor 1 and cellular inflammatory responses. Pharmacologically inhibiting autophagy using 3-methyladenine, a known inhibitor of class III phosphatidylinositol 3-kinase, confirms these effects. Either Atg5(-/-) or Atg7(-/-) MEFs are, independent of changes in IFN-gamma receptor expression, resistant to IFN-gamma-activated Jak2-STAT1, which suggests that autophagy is important for IFN-gamma signal transduction. Lentivirus-based short hairpin RNA for Atg5 knockdown confirmed the importance of autophagy for IFN-gamma-activated STAT1. Without autophagy, reactive oxygen species increase and cause SHP2 (Src homology-2 domain-containing phosphatase 2)-regulated STAT1 inactivation. Inhibiting SHP2 reversed both cellular inflammation and the IFN-gamma-induced activation of STAT1 in Atg5(-/-) MEFs. Our study provides evidence that there is a link between autophagy and both IFN-gamma signaling and cellular inflammation and that autophagy, because it inhibits the expression of reactive oxygen species and SHP2, is pivotal for Jak2-STAT1 activation.
MeSH Terms
Adenine/analogs & derivatives,pharmacology
Animals
Autophagy/physiology
Autophagy-Related Protein 5
Autophagy-Related Protein 7
Cells, Cultured
Embryo, Mammalian/cytology,metabolism
Enzyme Inhibitors/pharmacology
Fibroblasts/cytology,metabolism
Gene Knockdown Techniques
Inflammation/genetics,metabolism
Interferon-gamma/genetics,metabolism
Janus Kinase 2/genetics,metabolism
Lentivirus
Mice
Microtubule-Associated Proteins/genetics,metabolism
Phosphatidylinositol 3-Kinases/genetics,metabolism
Phosphoinositide-3 Kinase Inhibitors
Protein Tyrosine Phosphatase, Non-Receptor Type 11/genetics,metabolism
Reactive Oxygen Species/metabolism
STAT1 Transcription Factor/genetics,metabolism
Signal Transduction/drug effects,physiology
Chemicals
Atg5 protein, mouse
Atg7 protein, mouse
Autophagy-Related Protein 5
Enzyme Inhibitors
Microtubule-Associated Proteins
Phosphoinositide-3 Kinase Inhibitors
Reactive Oxygen Species
STAT1 Transcription Factor
Stat1 protein, mouse
3-methyladenine
Interferon-gamma
Jak2 protein, mouse
Janus Kinase 2
Protein Tyrosine Phosphatase, Non-Receptor Type 11
Ptpn11 protein, mouse
Autophagy-Related Protein 7
Adenine
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Chang Yu-Ping
Institutes of Clinical Medicine, College of Medicine, National Cheng Kung University, Tainan 701, Taiwan.
Tsai Cheng-Chieh
Huang Wei-Ching
Wang Chi-Yun
Chen Chia-Ling
Lin Yee-Shin
Kai Jui-In
Hsieh Chia-Yuan
Cheng Yi-Lin
Choi Pui-Ching
Chen Shun-Hua
Chang Shih-Ping
Liu Hsiao-Sheng
Lin Chiou-Feng
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