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Related Concept Videos

Chromatin Modification in iPS Cells01:32

Chromatin Modification in iPS Cells

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Chromatin modification alters gene expression; therefore, scientists can add histone-modifying enzymes, histone variants, and chromatin remodeling complexes to somatic cells to aid reprogramming into pluripotent stem (iPS) cells.
Compact chromatin makes reprogramming difficult. Enzymes, such as histone demethylases and acetyltransferases, are often added during reprogramming to loosen the chromatin, making the DNA more accessible to transcription factors. Molecules that inhibit histone...
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The histone proteins have a flexible N-terminal tail extending out from the nucleosome. These histone tails are often subjected to post-translational modifications such as acetylation, methylation, phosphorylation, and ubiquitination. Particular combinations of these modifications form “histone codes” that influence the chromatin folding and tissue-specific gene expression.
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Related Experiment Video

Updated: Apr 10, 2026

Complete Workflow for Analysis of Histone Post-translational Modifications Using Bottom-up Mass Spectrometry: From Histone Extraction to Data Analysis
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Fluctuations in histone H4 isoforms during cellular reprogramming monitored by middle-down proteomics.

Marco Benevento1,2, Peter D Tonge3, Mira C Puri3,4

  • 1Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.

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Summary

Cellular reprogramming involves epigenetic changes beyond histone H3. This study identifies key histone H4 modifications during induced pluripotency, revealing crucial roles in cellular reprogramming.

Keywords:
Cell biologyEpigeneticsHistonesMiddle-downPluripotencyReprogramming

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Area of Science:

  • Epigenetics and Molecular Biology
  • Cellular Biology
  • Proteomics

Background:

  • Cellular reprogramming resets somatic cells to a pluripotent state, involving epigenetic modifications.
  • Histone post-translational modifications are critical for this epigenetic remodeling.
  • Previous studies focused on histone H3 marks, leaving histone H4 roles less explored.

Purpose of the Study:

  • To comprehensively identify and quantify histone H4 isoforms and their modifications during cellular reprogramming.
  • To investigate the role of histone H4 modifications in establishing induced pluripotency.
  • To compare H4 modification patterns between somatic cells, reprogramming cells, and pluripotent cells (ESCs and iPSCs).

Main Methods:

  • Unbiased middle-down proteomics approach.
  • ChIP-sequencing (ChIP-seq) for histone H3 marks (from previous work).
  • Mass spectrometry (MS) for histone H4 isoform identification and quantification.

Main Results:

  • Identified 72 unique histone H4 isoforms and quantified 56.
  • Detected significant differences in H4 modifications between somatic and late-phase reprogramming cells.
  • Observed higher H4 acetylation and tri-methylation, with lower mono- and di-methylation in ESCs and iPSCs compared to reprogramming cells.

Conclusions:

  • Epigenetic remodeling during cellular reprogramming extends to histone H4 modifications.
  • Histone H4 modifications are important for the acquisition of induced pluripotency.
  • The findings provide a valuable resource for studying epigenetic mechanisms in pluripotency.