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Recognition of cytosolic DNA attenuates glucose metabolism and induces AMPK mediated energy stress response.

Zheng M, Xie L, Liang Y, Wu S, Xu H, Zhang Y, Liu H, Lin D, Han J, Lu K - Int. J. Biol. Sci. (2015)

Bottom Line: Recognition of cytosolic DNA activates a series of cellular responses, including induction of pro-inflammatory genes such as type I interferon through the well-known cGAS-STING pathway.Here we show for the first time that intracellular administration of either single or double stranded interferon stimulating DNA (ISD), but not poly(dA) suppresses cell growth in many different cell types.Suppression of cell growth by cytosolic DNA is cGAS/STING independent and associated with inhibition of glucose metabolism, ATP depletion and subsequent cellular energy stress responses including activation of AMPK and inactivation of mTORC1.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian, 350108, China. ; 2. Translational Medicine Institute, Fujian Medical University, Fuzhou, Fujian, 350108, China.

ABSTRACT
Both viral infection and DNA transfection expose single-stranded or double-stranded DNA to the cytoplasm of mammalian cells. Recognition of cytosolic DNA activates a series of cellular responses, including induction of pro-inflammatory genes such as type I interferon through the well-known cGAS-STING pathway. Here we show for the first time that intracellular administration of either single or double stranded interferon stimulating DNA (ISD), but not poly(dA) suppresses cell growth in many different cell types. Suppression of cell growth by cytosolic DNA is cGAS/STING independent and associated with inhibition of glucose metabolism, ATP depletion and subsequent cellular energy stress responses including activation of AMPK and inactivation of mTORC1. Our results suggest that in concert with but independent of innate immune response, recognition of cytosolic DNA induced cellular energy stress potentially functions as a metabolic barrier to viral replication.

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Related in: MedlinePlus

Cytosolic ISD induced metabolic stress is independent of cGAS and STING. (A) cGAS or STING were depleted using TALEN genome editing techniques. Cells were treated as indicated. WST-1 assay was performed 6 hours after transfection. (B) L929-cGAS-KO and L929-STING-KO were generated using TALEN genome editing techniques. Cells were then transfected with dsDNA for indicated time. IFNbeta mRNA was measured by quantitative RT-PCR. (C) Expression of STING was determined by anti-STING antibody. F: sense strand. R: anti-sense strand. Data of three independent replicates are presented as the mean +/- s.e.m., n=3.
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Figure 4: Cytosolic ISD induced metabolic stress is independent of cGAS and STING. (A) cGAS or STING were depleted using TALEN genome editing techniques. Cells were treated as indicated. WST-1 assay was performed 6 hours after transfection. (B) L929-cGAS-KO and L929-STING-KO were generated using TALEN genome editing techniques. Cells were then transfected with dsDNA for indicated time. IFNbeta mRNA was measured by quantitative RT-PCR. (C) Expression of STING was determined by anti-STING antibody. F: sense strand. R: anti-sense strand. Data of three independent replicates are presented as the mean +/- s.e.m., n=3.

Mentions: Due to the pivotal role of cGAS and STING in recognition of cytosolic DNA and induction of type I interferon 22, we examined whether cGAS or STING was required for ISD to induce metabolic stress. Because both cGAS and STING were known to express at very low levels in 293T cells 21, we used mouse fibroblast cells L929 as a model to study the relevance between cellular metabolic stress response and innate immune response, and their dependence on cGAS and STING. As shown in Fig 4, neither cGAS nor STING knockout attenuated the ability of either ssISD or dsISD induced metabolic stress response in L929 cells (Fig. 4A), although knockout of either cGAS or STING completely abolished cytosolic DNA induced type I interferon response (Fig. 4B and 4C), as expected12, 21. These results suggest that cytosolic ISD induced metabolic response is independent of cGAS and STING, and irrelevant to type I interferon mediated innate immune response.


Recognition of cytosolic DNA attenuates glucose metabolism and induces AMPK mediated energy stress response.

Zheng M, Xie L, Liang Y, Wu S, Xu H, Zhang Y, Liu H, Lin D, Han J, Lu K - Int. J. Biol. Sci. (2015)

Cytosolic ISD induced metabolic stress is independent of cGAS and STING. (A) cGAS or STING were depleted using TALEN genome editing techniques. Cells were treated as indicated. WST-1 assay was performed 6 hours after transfection. (B) L929-cGAS-KO and L929-STING-KO were generated using TALEN genome editing techniques. Cells were then transfected with dsDNA for indicated time. IFNbeta mRNA was measured by quantitative RT-PCR. (C) Expression of STING was determined by anti-STING antibody. F: sense strand. R: anti-sense strand. Data of three independent replicates are presented as the mean +/- s.e.m., n=3.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4400389&req=5

Figure 4: Cytosolic ISD induced metabolic stress is independent of cGAS and STING. (A) cGAS or STING were depleted using TALEN genome editing techniques. Cells were treated as indicated. WST-1 assay was performed 6 hours after transfection. (B) L929-cGAS-KO and L929-STING-KO were generated using TALEN genome editing techniques. Cells were then transfected with dsDNA for indicated time. IFNbeta mRNA was measured by quantitative RT-PCR. (C) Expression of STING was determined by anti-STING antibody. F: sense strand. R: anti-sense strand. Data of three independent replicates are presented as the mean +/- s.e.m., n=3.
Mentions: Due to the pivotal role of cGAS and STING in recognition of cytosolic DNA and induction of type I interferon 22, we examined whether cGAS or STING was required for ISD to induce metabolic stress. Because both cGAS and STING were known to express at very low levels in 293T cells 21, we used mouse fibroblast cells L929 as a model to study the relevance between cellular metabolic stress response and innate immune response, and their dependence on cGAS and STING. As shown in Fig 4, neither cGAS nor STING knockout attenuated the ability of either ssISD or dsISD induced metabolic stress response in L929 cells (Fig. 4A), although knockout of either cGAS or STING completely abolished cytosolic DNA induced type I interferon response (Fig. 4B and 4C), as expected12, 21. These results suggest that cytosolic ISD induced metabolic response is independent of cGAS and STING, and irrelevant to type I interferon mediated innate immune response.

Bottom Line: Recognition of cytosolic DNA activates a series of cellular responses, including induction of pro-inflammatory genes such as type I interferon through the well-known cGAS-STING pathway.Here we show for the first time that intracellular administration of either single or double stranded interferon stimulating DNA (ISD), but not poly(dA) suppresses cell growth in many different cell types.Suppression of cell growth by cytosolic DNA is cGAS/STING independent and associated with inhibition of glucose metabolism, ATP depletion and subsequent cellular energy stress responses including activation of AMPK and inactivation of mTORC1.

View Article: PubMed Central - PubMed

Affiliation: 1. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, Fujian, 350108, China. ; 2. Translational Medicine Institute, Fujian Medical University, Fuzhou, Fujian, 350108, China.

ABSTRACT
Both viral infection and DNA transfection expose single-stranded or double-stranded DNA to the cytoplasm of mammalian cells. Recognition of cytosolic DNA activates a series of cellular responses, including induction of pro-inflammatory genes such as type I interferon through the well-known cGAS-STING pathway. Here we show for the first time that intracellular administration of either single or double stranded interferon stimulating DNA (ISD), but not poly(dA) suppresses cell growth in many different cell types. Suppression of cell growth by cytosolic DNA is cGAS/STING independent and associated with inhibition of glucose metabolism, ATP depletion and subsequent cellular energy stress responses including activation of AMPK and inactivation of mTORC1. Our results suggest that in concert with but independent of innate immune response, recognition of cytosolic DNA induced cellular energy stress potentially functions as a metabolic barrier to viral replication.

Show MeSH
Related in: MedlinePlus