Chromosome Research

, Volume 17, Issue 1, pp 47–64 | Cite as

Chromatin condensation in terminally differentiating mouse erythroblasts does not involve special architectural proteins but depends on histone deacetylation

  • Evgenya Y. Popova
  • Sharon Wald Krauss
  • Sarah A. Short
  • Gloria Lee
  • Jonathan Villalobos
  • Joan Etzell
  • Mark J. Koury
  • Paul A. Ney
  • Joel Anne Chasis
  • Sergei A. Grigoryev
Article
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Abstract

Terminal erythroid differentiation in vertebrates is characterized by progressive heterochromatin formation and chromatin condensation and, in mammals, culminates in nuclear extrusion. To date, although mechanisms regulating avian erythroid chromatin condensation have been identified, little is known regarding this process during mammalian erythropoiesis. To elucidate the molecular basis for mammalian erythroblast chromatin condensation, we used Friend virus-infected murine spleen erythroblasts that undergo terminal differentiation in vitro. Chromatin isolated from early and late-stage erythroblasts had similar levels of linker and core histones, only a slight difference in nucleosome repeats, and no significant accumulation of known developmentally regulated architectural chromatin proteins. However, histone H3(K9) dimethylation markedly increased while histone H4(K12) acetylation dramatically decreased and became segregated from the histone methylation as chromatin condensed. One histone deacetylase, HDAC5, was significantly upregulated during the terminal stages of Friend virus-infected erythroblast differentiation. Treatment with histone deacetylase inhibitor, trichostatin A, blocked both chromatin condensation and nuclear extrusion. Based on our data, we propose a model for a unique mechanism in which extensive histone deacetylation at pericentromeric heterochromatin mediates heterochromatin condensation in vertebrate erythroblasts that would otherwise be mediated by developmentally-regulated architectural proteins in nucleated blood cells.

Key words

erythroblast enucleation chromatin condensation heterochromatin histone deacetylation 

Abbreviations

FVA cells

Friend virus-infected murine spleen erythroblasts

H3acK9,K14

histone H3 acetylated at lysines 9 and 14

H3me2K9

histone H3 dimethylated at lysine 9

H3me3K9

histone H3 trimethylated at lysine 9

H4acK12

histone H4 acetylated at lysine 12

HDAC

histone deacetylase

HP1

heterochromatin protein 1

HPLC

high-performance liquid chromatography

KCM buffer

120 mM KCl, 20 mM NaCl, 10 mM Tris-HCl, pH 7.7, 0.1% Triton X-100

MEL

mouse erythroleukemia cells

PBS

phosphate-buffered saline

RSB

reticulocyte standard buffer (10 mM NaCl, 3 mM MgCl2, 10 mM Hepes, pH 7.5)

SDS-PAGE

sodium dodecyl sulfate–polyacrylamide gel electrophoresis

TSA

trichostatin A

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Notes

Acknowledgments

We are grateful to Drs. P. Singh (Borstel, Germany), D. Tremethick (Canberra, Australia), and N. Chaudhary (Woodlands, TX, USA) for their kind gifts of antibodies against HP1, H2A.Z, and lamins A/C and B. Supported in part by National Institutes of Health Grants DK32094, DK56267, DK59079, and CA084214; National Science Foundation grant MCB-0615536, and by the Director, Office of Health and Environment Research Division, US Department of Energy, under Contract DE-AC03-76SF00098; and by Merit Review Award from the Department of Veteran Affairs.

Supplementary material

10577_2008_9005_MOESM1_ESM.doc (32 kb)
Tab1 (DOC 32.5 kb)

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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Evgenya Y. Popova
    • 1
  • Sharon Wald Krauss
    • 2
  • Sarah A. Short
    • 2
  • Gloria Lee
    • 2
  • Jonathan Villalobos
    • 2
  • Joan Etzell
    • 3
  • Mark J. Koury
    • 4
  • Paul A. Ney
    • 5
  • Joel Anne Chasis
    • 2
  • Sergei A. Grigoryev
    • 1
  1. 1.Biochemistry and Molecular Biology, College of MedicinePenn State UniversityHersheyUSA
  2. 2.Life Sciences DivisionUniversity of California, Lawrence Berkeley National LaboratoryBerkeleyUSA
  3. 3.Laboratory MedicineUniversity of CaliforniaSan FranciscoUSA
  4. 4.Medicine/HematologyTN Valley Healthcare System and Vanderbilt UniversityNashvilleUSA
  5. 5.St. Jude Children’s Research HospitalMemphisUSA

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