Involvement of GATA1 and Sp3 in the activation of the murine STING gene promoter in NIH3T3 cells

Scientific Reports
Yan-Yan XuHua-Guo Xu

Abstract

Stimulator of Interferon Gene (STING) is a key mediator of innate immune signaling. STING plays a pivotal role in the pathogenesis of many diseases including infectious diseases, auto-immune diseases and cancer. Many studies have been carried out recently in the field of STING-regulated pathway, however, rarely of transcriptional mechanisms. To characterize the murine STING (mSTING) promoter, we cloned a series of different nucleotide sequences of the 5'-flanking region of the mSTING gene. Transient transfection of promoter-reporter recombinant plasmids and luciferase assay illustrated the region (-77/+177) relative to the transcription start site (TSS) of the mSTING gene was sufficient for full promoter activity. This region contains GATA1, IK2, Sp1/Sp3 and STAT putative transcription factor binding sites. Mutation of GATA1 or Sp1/Sp3 sites led to obvious decrease of the mSTING promoter activity. Overexpression of GATA1 and Sp3 enhanced the mSTING promoter activity, whereas knockdown of GATA1 and Sp3 by a siRNA strategy significantly reduced the transcription activity. Chromatin immunoprecipitation assays demonstrated that GATA1 and Sp3 interact with the mSTING promoter in vivo. These results provided the first analysis of mST...Continue Reading

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Citations

Oct 28, 2020·PLoS Pathogens·Mike KhanJudith A Smith

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Methods Mentioned

BETA
immunoprecipitation
PCR
transfections
transfection
ChIP
protein assay
electrophoresis

Software Mentioned

TFSEARCH
Primer Primier
SPSS
ENCODE

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