Dataset Information

TgCoq5 proximity proteome in mitochondria and cytosolic fractions using FbxO14 as spatial reference

ABSTRACT: Coenzyme Q or Ubiquinone (UQ) is an essential electron carrier and antioxidant. In yeast and mammals, several enzymes involved in the UQ synthesis form a protein complex in the mitochondrial matrix. Blast searches of the Toxoplasma gondii proteome failed to reveal several essential enzymes of the ubiquinone synthesis pathway. With the aim to identify those missing enzymes, we generated mutant strain expressing TurboID at the C-terminus of TgCoq5, one of the enzymes we have validated to be involved in UQ synthesis in T. gondii. Using a strategy combining biotinylation of TgCoq5-TurboID and FbxO14-TurboID, used as spatial reference, and subcellular fractionation. We investigated the proximal proteome of TgCoq5 and FbxO14 in the mitochondrial enriched fractions, P2, and cytosolic enriched fractions, S4.

INSTRUMENT(S):

ORGANISM(S): Toxoplasma Gondii Gt1

SUBMITTER: Baihetiya Baierna

LAB HEAD: Silvia Moreno

PROVIDER: PXD059833 | Pride | 2025-07-31

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

Items per page:

1 - 5 of 16

Similar Datasets

Project description:Circadian rhythms are daily oscillations in metabolism and physiology and are generated by the circadian clock. In fruit fly Drosophila, the circadian clock is generated by a transcription-translation feedback loop in which the positive arm components Clock and Cycle activate the expression of the Period and Timeless genes of negative arm, as well as other circadian clock-regulated genes. After being retained in the cytoplasm, the Period and Timeless proteins then migrate to the nucleus to inhibit the Clock/Cycle transactivity by protein-protein interactions (PPIs). The endogenous circadian clock is synchronized with the geological (solar) clock via photoreceptors. Drosophila Cryptochrome protein functions as a circadian photoreceptor. In the early morning, exposure of Cryptochrome to light causes a conformational change in it which results in the formation of new PPIs. Light-activated Cryptochrome interacts with the core clock protein Timeless and the E3 ubiquitin ligase-substrate adaptor protein Jetlag, which results in the ubiquitylation of Timeless by Jetlag-E3 ligase complex and then degradation of Timeless within minutes by proteasome system. Rapid degradation of Timeless and then its partner protein Period, because of its instability in the absence of Timeless, relieves the inhibition on the Clock/Cycle transcription factors suddenly. Therefore, Clock/Cycle-driven expression of circadian clock-regulated genes are induced again, which is the restart of the circadian oscillation or the resetting of the clock. Following Timeless degradation, Cryptochrome is also degraded so the photoreceptor mechanism does not start a new resetting signal until all the required factors are re-synthesized in a circadian manner. Light-dependent degradation of Drosophila Cryptochrome can be observed in Drosophila S2 cell line in culture. In this project, we aimed at finding the interactome of Cryptochrome protein in Drosophila S2 cell line under light and in the dark using proximity labeling method. Because of the fast kinetics of Cryptochrome degradation, we chose the enzymes that can saturate in less than one hour. TurboID (TID) and APEX2 enzymes label proteins with biotin in the proximity even though they work with different mechanisms. They were fused to Cryptochrome protein, and proximity labeling was performed in the dark or under light. We have identified novel light-dependent or -independent interactors of Drosophila Cryptochrome and confirmed some of them using classical coimmunoprecipitation technique.

			Action	DRS
	5P21.dat	Other
	5P21.raw	Raw
	5P22.dat	Other
	5P22.raw	Raw
	5S41.dat	Other

Dataset Information

TgCoq5 proximity proteome in mitochondria and cytosolic fractions using FbxO14 as spatial reference

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets