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Chromosome Research

, Volume 24, Issue 2, pp 161–173 | Cite as

RbAp48 is essential for viability of vertebrate cells and plays a role in chromosome stability

  • Pasjan Satrimafitrah
  • Hirak Kumar Barman
  • Ahyar Ahmad
  • Hideki Nishitoh
  • Tatsuo Nakayama
  • Tatsuo Fukagawa
  • Yasunari TakamiEmail author
Article

Abstract

RbAp46/48, histone chaperone, is a family of evolutionarily conserved WD40 repeat-containing proteins, which are involved in various chromatin-metabolizing processes, but their in vivo functional relevance is yet unclear. In order to examine the biological role of pRbAp48 in chicken DT40 cells, we generated a tetracycline-inducible system for conditional RbAp48-knockout cells. Depletion of RbAp48 led to delayed S phase progression associated with slow DNA synthesis and nascent nucleosome formation, followed by accumulation in G2/M phase, finally leading to cell death. Prior to cell death, these cells exhibited aberrant mitosis such as highly condensed and abnormal chromosome alignment on the metaphase plate, leading to chromosome missegregation. Depletion of RbAp48 also caused dissociation of heterochromatin protein 1 (HP1) from pericentromeric heterochromatin. Furthermore, depletion of RbAp48 from cells led to elevated levels of acetylation and slightly decreased levels of methylation, specifically at Lys-9 residue of histone H3. These results suggest that RbAp48 plays an important role in chromosome stability for proper organization of heterochromatin structure through the regulation of epigenetic mark.

Keywords

Histone chaperone Chromatin assembly Histone modification WD40 protein Heterochromatin 

Abbreviations

CAF-1

Chromatin assembly factor

HP1

Heterochromatin protein 1

Rb

Retinoblastoma

HAT

Histone-acetyltransferase

HDAC

Histone deacetylase

HMT

Histone methyltransferase

Notes

Acknowledgments

This work was supported in part by Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan. We thank Ryoko Masuya and Nahoko Nagamatsu-Yamamoto for their technical assistance and Radha Madhyastha and HarishKumar Madhyastha for useful comments and critical reading of the manuscript.

Compliance with ethical standards

Conflict of interest

All the authors declare that they have no conflict of interest of any sort with anyone.

Supplementary material

10577_2015_9510_MOESM1_ESM.avi (2.1 mb)
Supplementary file 1 The live-cell observation was started at 36 h after plating the conditional RbAp48-knockout cells, endogenously tagged with DsRed and EGFP in the normal condition (without tet), and time-lapse images were taken every 20 min for the next 23 h. Time is shown as hours:minutes after addition of tet. Left panel, phase contrast; center panel, CENPH-GFP; right panel, HP1γ–DsRed. (AVI 2162 kb)
10577_2015_9510_MOESM2_ESM.avi (2.3 mb)
Supplementary file 2 The live-cell observation was started at 36 h after plating the conditional RbAp48-knockout cells, endogenously tagged with DsRed and EGFP in the presence of tet, and time-lapse images were taken every 20 min for the next 23 h. Time is shown as hours:minutes after addition of tet. Left panel, phase contrast; center panel, CENPH-GFP; right panel, HP1γ–DsRed. (AVI 2345 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Pasjan Satrimafitrah
    • 1
  • Hirak Kumar Barman
    • 1
    • 3
  • Ahyar Ahmad
    • 1
    • 4
  • Hideki Nishitoh
    • 1
  • Tatsuo Nakayama
    • 1
  • Tatsuo Fukagawa
    • 2
  • Yasunari Takami
    • 1
    Email author
  1. 1.Section of Biochemistry and Molecular Biology, Department of Medical SciencesUniversity of MiyazakiKiyotakeJapan
  2. 2.Department of Molecular GeneticsNational Institute of GeneticsMishimaJapan
  3. 3.Fish Genetics & Biotechnology Division, Central Institute of Freshwater AquacultureIndian Council of Agricultural ResearchBhubaneswarIndia
  4. 4.Department of ChemistryHasanuddin UniversityMakassarIndonesia

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