Dataset Information

Phosphoproteome in mitotic chromosomes

ABSTRACT: Identification and characterization of mitosis-specific phosphorylation in mitotic chromosome-associated proteins

INSTRUMENT(S): Q Exactive

ORGANISM(S): Gallus Gallus (chicken)

TISSUE(S): B Cell

SUBMITTER: Shinya Ohta

LAB HEAD: Shinya Ohta

PROVIDER: PXD003877 | Pride | 2016-09-27

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	11file-merge.zip	Other
	11files-mergemascot.txt.zip	Txt
	131212st2-DrOhta-sample_Phospho-400ug.raw	Raw
	131213st2-DrOhta-sample_Phospho-100ug.raw	Raw
	131213st2-DrOhta-sample_Phospho-200ug.raw	Raw

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Publications

Identification of Mitosis-Specific Phosphorylation in Mitotic Chromosome-Associated Proteins.

Ohta Shinya S Kimura Michiko M Takagi Shunsuke S Toramoto Iyo I Ishihama Yasushi Y

Journal of proteome research 20160817 9

During mitosis, phosphorylation of chromosome-associated proteins is a key regulatory mechanism. Mass spectrometry has been successfully applied to determine the complete protein composition of mitotic chromosomes, but not to identify post-translational modifications. Here, we quantitatively compared the phosphoproteome of isolated mitotic chromosomes with that of chromosomes in nonsynchronized cells. We identified 4274 total phosphorylation sites and 350 mitosis-specific phosphorylation sites i ...[more]

PMID: 27504668

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Project description:Mitosis entails global alterations to chromosome structure and nuclear architecture, concomitant with transient silencing of transcription. How cells transmit transcriptional states through mitosis remains incompletely understood. While many nuclear factors dissociate from mitotic chromosomes, the observation that certain nuclear factors and chromatin features remain associated with individual loci during mitosis originated the hypothesis that they could provide transcriptional memory through mitosis. To obtain the first genome-wide view of the dynamics of chromatin structure during mitosis, we compared the DNase sensitivity of interphase and mitotic chromatin at two stages of cellular maturation in a rapidly dividingmurine erythroblastmodel. Despite global chromosome condensation visible during mitosis at the microscopic level, the chromatin accessibility landscape is largely unaltered. However, mitotic chromatin accessibility is locally dynamic, with individual loci maintaining none, some, or all of their interphase accessibility. Mitotic reduction in accessibility occurs primarily within narrow, highly hypersensitive sites that frequently coincide with transcription factor binding sites, whereas broader domains of moderate accessibility tend to be more stable. In mitosis, proximal promoters generally maintain their accessibility, whereas distal regulatory elements preferentially lose accessibility. Promoters with the highest degree of accessibility preservation in mitosis tend to also be accessible across many murine tissues in interphase. Transcription factor GATA1 exerts site-specific changes in interphase accessibility that are most pronounced at distal regulatory elements, but does not visibly influence mitotic accessibility. We conclude that features of open chromatin are remarkably stable through mitosis and are modulated at the level of individual genes and regulatory elements. Dnase-Seq data is integrated with Chip-seq [GSE36589, GSE30142] and RNA-seq to examine epigentic changes in mitosis. We performed DNase-seq on two cell lines, G1E and G1E-ER4, both on an asynchronus population, and on a sample of cells in mitosis; each of the 4 experiments in triplicate.