Dataset Information

Crosstalk between Drp1 phosphorylations during mitochondrial remodelling and their impact on metabolic adaptation - Parallel reaction monitoring (PRM) mass spectrometry-based proteomic data

ABSTRACT: Mitochondria constantly undergo fusion and fission events, referred as mitochondrial dynamics, which determine intracellular mitochondrial architecture and bioenergetics. Cultured cell studies demonstrate that mitochondrial dynamics are acutely regulated by phosphorylation of the mitochondrial fission orchestrator Drp1, at S579 or S600. This dataset is linked to a study reporting on how in differentiated mouse tissues Drp1 phosphorylation at S600 acted as an upstream event for S579 phosphorylation, leading to mitochondrial fragmentation. We performed liquid chromatography mass spectrometry (LC-MS) analyses in phospho-Drp1 immunoprecipitates from BAT of vehicle or CL316,243-treated mice, following digestion with endoproteinase Glu-C to assess whether S600 and S579 phosphorylations were both occurring in the same Drp1 proteoforms or individually in different pools of Drp1 proteins. Our results certify that upon PKA stimulation, S600 and S579 phosphorylations occurred simultaneously on the same Drp1 molecular form.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Brown Adipose Tissue

SUBMITTER: Loïc Dayon

LAB HEAD: Loïc Dayon

PROVIDER: PXD027108 | Pride | 2021-08-26

REPOSITORIES: Pride

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Project description:Purpose:to verify Drp1-mediated biological pathways under physiological and ischemia conditions.Methods:C57BL/6 mice and Drp1 knockdown mice were used for ischemia treatment. Right femoral arteries were catheterized with polyethylene catheters for bleeding 50% of total blood volume (≈7% of weight). The ischemia time started to calculate after the model was established. A laparotomy was then carried out to obtain superior mesenteric artery tissues (SMAs) for transcriptome analysis. Series-Cluster analysis was performed to identify the global trends of mitochondrial DEGs after Drp1 knockdown. Gene ontology (GO) analysis was performed to facilitate elucidating the biological process (BP), molecular function (MF) and cellular component (CC) of unique genes in the significant or representative profiles. Results:In addition to its traditional roles in GTPase regulation and mitochondrial fission, Drp1 may also regulate actin cytoskeleton and the movement of microtubules. Besides, Drp1 may participate in the regulation of morphology and functions of other organelles, including endoplasmic reticulum, peroxisome, phagolysosome, etc. As for general organ functions, Drp1 may regulate vascular constriction and dilation. These consequences indicated the important role of Drp1 in ischemia-induced cellular regulation. Conclusions: Drp1 deficiency proves the important role of Drp1 in ischemia-induced biomedical processes through multiple potential fission-independent manners. Methods:C57BL/6 mice and Drp1 knockdown mice were used for ischemia treatment. Right femoral arteries were catheterized with polyethylene catheters for bleeding 50% of total blood volume (≈7% of weight). The ischemia time started to calculate after the model was established. A laparotomy was then carried out to obtain superior mesenteric artery tissues (SMAs) for transcriptome analysis. Series-Cluster analysis was performed to identify the global trends of mitochondrial DEGs after Drp1 knockdown. Gene ontology (GO) analysis was performed to facilitate elucidating the biological process (BP), molecular function (MF) and cellular component (CC) of unique genes in the significant or representative profiles. Results:In addition to its traditional roles in GTPase regulation and mitochondrial fission, Drp1 may also regulate actin cytoskeleton and the movement of microtubules. Besides, Drp1 may participate in the regulation of morphology and functions of other organelles, including endoplasmic reticulum, peroxisome, phagolysosome, etc. As for general organ functions, Drp1 may regulate vascular constriction and dilation. These consequences indicated the important role of Drp1 in ischemia-induced cellular regulation. Conclusions: Drp1 deficiency proves the important role of Drp1 in ischemia-induced biomedical processes through multiple potential fission-independent manners.

Dataset Information

Crosstalk between Drp1 phosphorylations during mitochondrial remodelling and their impact on metabolic adaptation - Parallel reaction monitoring (PRM) mass spectrometry-based proteomic data

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