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H2B variants expression during iAs-mediated EMT

ABSTRACT: This work details the first global mass spectrometric analysis of histone H2B variants as cells undergo arsenic-mediated epithelial to mesenchymal transition.

INSTRUMENT(S): Fourier Transform Ion Cyclotron Resonance Mass Spectrometer

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Lung, Hela Cell

SUBMITTER: Tingting Jiang

LAB HEAD: Yvonne N. Fondufe-Mittendorf

PROVIDER: PXD003503 | Pride | 2016-05-17

REPOSITORIES: Pride

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			Action	DRS
	Beas_NT_Broadband_100scans_rep1.dat	Other
	Beas_NT_Broadband_100scans_rep2.dat	Other
	Beas_NT_Broadband_100scans_rep3.dat	Other
	Beas_T_Broadband_100scans_rep1.dat	Other
	Beas_T_Broadband_100scans_rep2.dat	Other

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Publications

Quantitative Mass Spectrometry Reveals Changes in Histone H2B Variants as Cells Undergo Inorganic Arsenic-Mediated Cellular Transformation.

Rea Matthew M Jiang Tingting T Eleazer Rebekah R Eckstein Meredith M Marshall Alan G AG Fondufe-Mittendorf Yvonne N YN

Molecular & cellular proteomics : MCP 20160511 7

Exposure to inorganic arsenic, a ubiquitous environmental toxic metalloid, leads to carcinogenesis. However, the mechanism is unknown. Several studies have shown that inorganic arsenic exposure alters specific gene expression patterns, possibly through alterations in chromatin structure. While most studies on understanding the mechanism of chromatin-mediated gene regulation have focused on histone post-translational modifications, the role of histone variants remains largely unknown. Incorporati ...[more]

PMID: 27169413

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Pancreatic Cancer HPAF/CD18 cells: Ecd knockdown vs Scrambled

Project description:Purpose: To study the expression and function of a novel cell cycle regulatory protein, human ecdysoneless (Ecd), during pancreatic cancer (PC) pathogenesis. Experimental Design: Immunohistochemical expression profiling of Ecd was done in non-neoplastic normal pancreatic tissues and pancreatic ductal adenocarcinoma lesions (from tissue microarray and Rapid Autopsy program) as well as precancerous PanIN lesions and metastatic organs. To analyze the biological significance of Ecd in PC progression, Ecd was stably knocked down in PC cell line followed by in vitro and in vivo functional assays. Results: Normal pancreatic ducts show very weak to no Ecd expression compared to significant positive expression in PC tissues (mean±SE composite score: 0.3±0.2 and 3.8±0.2 respectively, p<0.0001) as well as in PanIN precursor lesions with a progressive increase in Ecd expression with increasing dysplasia (PanIN-1 to PanIN-3). Analysis of matched primary tumors and metastases from PC patients revealed that Ecd is highly expressed in both primary pancreatic tumor and in distant metastatic sites. Further, knockdown of Ecd suppressed cell proliferation in vitro and tumorigenicity of PC cells in mice orthotopic tumors. Microarray study revealed that Ecd regulates expression of glucose transporter GLUT4 in PC cells and was subsequently shown to modulate glucose uptake, lactate production and ATP generation by PC cells. Finally, knockdown of Ecd also reduced level of pAkt, key signaling molecule known to regulate aerobic glycolysis in cancer cells. Conclusion: Ecd is a novel tumor promoting factor that is differentially expressed in pancreatic cancer and potentially regulates glucose metabolism within cancer cells. Two-condition experiment, Ecd knockdown vs Scrambled cells. Biological replicates: 3 Ecd knockdownl, 3 Scrambled, independently grown and harvested. One replicate per array

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Systematic quantitative analysis of H2A and H2B variants by targeted proteomics

Project description:Background: Histones organize DNA into chromatin through a variety of processes. Among them, a vast diversity of histone variants can be incorporated into chromatin to finely modulate its organization and functionality. Classically, the study of histone variants has largely relied on antibody-based assays. However, antibodies have a limited efficiency to discriminate between highly similar histone variants. Results: In this study, we established a mass spectrometry-based analysis to address this challenge. We developed a targeted proteomics method, using Single Reaction Monitoring (SRM) or Parallel Reaction Monitoring (PRM), to quantify a maximum number of histone variants in a single multiplexed assay, even when histones are present in a crude extract. This strategy was developed on H2A and H2B variants, using 55 peptides corresponding to 25 different histone sequences, among which a few differ by a single amino acid. The methodology was then applied to mouse testis extracts in which almost all histone variants are expressed. It confirmed the abundance profiles of several testis-specific histones during successive stages of spermatogenesis and the existence of predicted H2A.L.1 isoforms. This methodology was also used to explore the over-expression pattern of H2A.L.1 isoforms in a mouse model of male infertility. Conclusions: Our results demonstrate that targeted proteomics is a powerful method to quantify highly similar histone variants and isoforms. The developed method can be easily transposed to the study of human histone variants, whose abundance can be deregulated in various diseases.

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Electron Capture vs. Transfer Dissociation for Site Determination of Tryptic Peptide Tyrosine Sulfation: Direct Detection of Fibrinogen Sulfation Sites and Identification of Novel Isobaric Interferences

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2024-09-05 | PXD052561 | Pride

Molecular basis of RanGTP-activated release of Histones H2A-H2B from Importin-9

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The Isotopic Depletion Increases the Spatial Resolution of FPOP Top-Down Analysis.

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Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

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Examination of histone H2B ubiquitination in wild type and USP49 knockdown cells [ChIP-Seq]

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