Project description:We examine the role of G3bp1, a RNA binding protein and site specific endoribonuclease in gene expression in isolated neonatal cardiomyocytes. RNAseq data from cardiomyocytes were infected with adenoviruses expressing shRNA against G3bp1 (ad-siG3bp1) or Luciferase (ad-siLUC, control) showed significant decrease in transcript abudnnace of cardiac-enriched genes involved in Calcium handling, contraction, action potential and sacromere function. On the other hand increase was observed in genes that regulate Hippo, TNF and TGFb signaling. Knockdown of G3bp1 inhibited endothelin-1 induced cardiomyocyte hypertrophy.
Project description:Mitochondrial calcium signaling plays a critical role in mitochondrial homeostasis during non-alcoholic steatohepatitis (NASH) progression. Here, we report Ras‐GTPase activating protein SH3 domain‐binding protein 1 (G3BP1), a core protein of stress granule (SG), significantly upregulated in NAFLD/NASH patients, mouse models and palmitic acid-stimulated hepatocytes. Hepatocyte-specific G3BP1 deficiency attenuates NASH in two dietary mouse models. SG and the mitochondrial import protein TOM70 collaboratively mediate the translocation of G3BP1 to the mitochondria. G3BP1 interacts and stabilizes mitochondrial calcium uptake 1 (MICU1) via inhibiting YME1L1-mediated degradation of MICU1, and also impedes MCU complex activity and assembly, leading to mitochondrial calcium overload. Moreover, metabolic stress suppresses TRIM25-mediated K63-ubiquitination of G3BP1 and subsequent SG disassembly. Pharmacological inhibition of G3BP1 impairs the G3BP1-MICU1 interaction and prevents mitochondrial homeostasis imbalance and NASH progression. Collectively, we uncover the significant role of mitochondrial G3BP1 in mitochondrial homeostasis and NASH progression, which provides a potential target for therapeutic interventions.
Project description:We previously identified the stimuli-responsive lncRNA CALA to impact endothelial cell function and identified G3BP1 as a main interaction partner of CALA in the cytoplasm of human umblical vein endothelial cells (HUVECs). To uncover the role of the lncRNA CALA in G3BP1-RNPs in the cytoplasm, we purified the G3BP1 interactome in control and CALA-silenced HUVECs via anti-G3BP1 immunoprecipitation followed by mass spectrometry. We thereby uncover the basal G3BP1 protein interactome in HUVECs and additionally identify lncRNA-dependent protein interactions of G3BP1 in the cytoplasm of HUVEC.
Project description:To comprehensively understand the characterization of bluetongue virus (BTV)-host interactome, and BTV infection and pathogenic mechanisms, RNA-seq was performed with BTV serotype 1 Y863 strain-infected and mock-infected sheep embryonic testicular cells (OA3.Ts) at 24 hours post-infection.
Project description:To comprehensively understand the characterization of bluetongue virus (BTV)-host interactome, and BTV infection and pathogenic mechanisms, RNA-seq was performed with BTV serotype 1 Y863 strain-infected and mock-infected sheep embryonic testicular cells (OA3.Ts) at 24 hours post-infection.
Project description:Analysis of U2AF2-TurboID-HA interactions by immunoprecipitation for comparison with proximity labeling. Cells expressing U2AF2 (iU2) fused with TurboID in either the undifferentiated state (D0) or after mesoderm differentiation (D1) were analyzed to identify the interactome and detect changes during the differentiation process.