Dataset Information

Diurnal regulatory landscape of the mouse liver nucleus

ABSTRACT: To dissect how diurnal rhythms affect key functions such as transcription or chromatin remodelling, we quantified the temporal nuclear accumulation of proteins and phosphoproteins from mouse liver using in vivo stable isotope labelling by amino acids in cell culture (SILAC)-based mass spectrometry (MS). Protein extracts from isotope labelled mice liver nuclei were used as a reference and mixed with extracts from animals collected every 3 h for 45 h total. Phosphopeptide levels were analysed after enrichment with titanium dioxide (TiO2). The proteins levels were also analysed (dataset with project accession PXD003818).

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Hepatocyte, Liver

SUBMITTER: Loïc Dayon

LAB HEAD: Frédéric Gachon

PROVIDER: PXD004191 | Pride | 2016-12-22

REPOSITORIES: Pride

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Publications

Nuclear Proteomics Uncovers Diurnal Regulatory Landscapes in Mouse Liver.

Wang Jingkui J Mauvoisin Daniel D Martin Eva E Atger Florian F Galindo Antonio Núñez AN Dayon Loïc L Sizzano Federico F Palini Alessio A Kussmann Martin M Waridel Patrice P Quadroni Manfredo M Dulić Vjekoslav V Naef Felix F Gachon Frédéric F

Cell metabolism 20161103 1

Diurnal oscillations of gene expression controlled by the circadian clock and its connected feeding rhythm enable organisms to coordinate their physiologies with daily environmental cycles. While available techniques yielded crucial insights into regulation at the transcriptional level, much less is known about temporally controlled functions within the nucleus and their regulation at the protein level. Here, we quantified the temporal nuclear accumulation of proteins and phosphoproteins from mo ...[more]

PMID: 27818260

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Project description:Being inspired by metabolomic data processing, we have developed a bioinformatic pipeline that optimizes the processing of mass spectral data obtained from isobaric Tandem Mass Tag (TMT) experiments. Our method focuses on the tandem mass spectral level by first quantifying and then identifying (QtI), while preserving unidentified spectra for further investigations. The raw datasets were previously generated [1, 2]. Two-proteome model experiments were considered where identical pools of human CSF or plasma samples were mixed with E. coli samples at different concentrations. E. coli protein extract was spiked in 400 µL of CSF at amounts of 0, 2, 3, 5, 6.25, and 7.5 µg. Such sets of 6 spiked CSF samples were prepared in triplicate for comparison using sixplex isobaric tagging and analyzed in triplicates on two independent but identical LC MS/MS, for a total of 18 raw files [1]; this experiment is called “CSF-E.coli”. E. coli protein extract was spiked at 0, 2.5, 5, 6.25, 12.5, and 25 µg in 30 µL human plasma. Such sets of 6 spiked plasma samples were prepared in quadruplicate for comparison using sixplex isobaric tagging and analyzed in triplicates on one LC MS/MS, for a total of 12 raw files [2]; this experiment is referred as “Plasma-E.coli”.The so-called “96samples-CSF” experiment consists of 16 replicate TMT sixplex experiments measuring identical CSF samples from the pool described above [2], analyzed in triplicates on one LCMS/MS for a total of 48 raw files. The so-called “96samples-plasma” experiment consists of 16 replicate TMT sixplex experiments measuring identical plasma samples from the pool described before, analyzed in triplicates on one LC MS/MS for a total of 48 raw files [1]. References: [1] Dayon, L., Núñez Galindo, A., Corthésy, J., Cominetti, O. & Kussmann, M. Comprehensive and scalable highly automated MS-based proteomic workflow for clinical biomarker discovery in human plasma. J. Proteome Res. 13, 3837-3845 (2014). [2] Núñez Galindo, A., Kussmann, M. & Dayon, L. Proteomics of Cerebrospinal Fluid: Throughput and Robustness Using a Scalable Automated Analysis Pipeline for Biomarker Discovery. Anal. Chem. 87, 10755-10761 (2015).

Dataset Information

Diurnal regulatory landscape of the mouse liver nucleus

Publications

Nuclear Proteomics Uncovers Diurnal Regulatory Landscapes in Mouse Liver.

Similar Datasets

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