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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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Intracellular localization is essential for ISD induced metabolic stress. (A) 293T Cells were treated as indicated. WST-1 assay was performed at 6 hours post-transfection. (B) Real-time live cell imaging was taken from 5 minutes to 4 hours after transfection with ssiSD labeled with Cy5. Pictures were taken every 5 minutes. Data of three independent replicates are presented as the mean +/- s.e.m., n=3.
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Figure 3: Intracellular localization is essential for ISD induced metabolic stress. (A) 293T Cells were treated as indicated. WST-1 assay was performed at 6 hours post-transfection. (B) Real-time live cell imaging was taken from 5 minutes to 4 hours after transfection with ssiSD labeled with Cy5. Pictures were taken every 5 minutes. Data of three independent replicates are presented as the mean +/- s.e.m., n=3.

Mentions: To determine whether this phenotype requires intracellular localization of DNA, we incubated ssISD or dsISD in the presence or absence of the transfection vehicle, which introduced DNA into cells. WST-1 reduction was decreased only in cells treated with the vehicle-DNA complex, but not ISD alone (Fig. 3A), indicating that recognition of ISD occurred in the cytoplasm, but not on the membrane. Furthermore, live-cell assay showed that Cy5 labeled ssISD entered cells as early as 2 hours post-transfection (Fig. 3B), which was consistent with the time point when WST-1 assay showed dramatic change. These data suggest that it is intracellular ISD that induces such phenotype, as shown for other ISD responses 21.


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)

Intracellular localization is essential for ISD induced metabolic stress. (A) 293T Cells were treated as indicated. WST-1 assay was performed at 6 hours post-transfection. (B) Real-time live cell imaging was taken from 5 minutes to 4 hours after transfection with ssiSD labeled with Cy5. Pictures were taken every 5 minutes. Data of three independent replicates are presented as the mean +/- s.e.m., n=3.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 3: Intracellular localization is essential for ISD induced metabolic stress. (A) 293T Cells were treated as indicated. WST-1 assay was performed at 6 hours post-transfection. (B) Real-time live cell imaging was taken from 5 minutes to 4 hours after transfection with ssiSD labeled with Cy5. Pictures were taken every 5 minutes. Data of three independent replicates are presented as the mean +/- s.e.m., n=3.
Mentions: To determine whether this phenotype requires intracellular localization of DNA, we incubated ssISD or dsISD in the presence or absence of the transfection vehicle, which introduced DNA into cells. WST-1 reduction was decreased only in cells treated with the vehicle-DNA complex, but not ISD alone (Fig. 3A), indicating that recognition of ISD occurred in the cytoplasm, but not on the membrane. Furthermore, live-cell assay showed that Cy5 labeled ssISD entered cells as early as 2 hours post-transfection (Fig. 3B), which was consistent with the time point when WST-1 assay showed dramatic change. These data suggest that it is intracellular ISD that induces such phenotype, as shown for other ISD responses 21.

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