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Chromosome Research

, Volume 20, Issue 3, pp 319–331 | Cite as

Roles of histone H3K9 methyltransferases during Drosophila spermatogenesis

  • Yuta Ushijima
  • Yoshihiro H. Inoue
  • Takahiro Konishi
  • Daishi Kitazawa
  • Hideki Yoshida
  • Kouhei Shimaji
  • Hiroshi Kimura
  • Masamitsu YamaguchiEmail author
Article

Abstract

Epigenetic regulation of gene expression by covalent modification of histones is important for germ line cell development. In mammals, histone H3 lysine 9 (H3K9)-specific histone methyltransferases (HMTases), such as G9a, SETDB1, and SUV39H, play critical roles, but the contribution of H3K9-specific HMTases in Drosophila remains to be clarified, especially in male sperm. Here, we performed immunocytochemical analyses with a specific antibody to dG9a, Drosophila G9a ortholog, and demonstrated localization in the cytoplasm from the growth to elongation stages of spermatogenesis. In the subsequent early canoe stage, strong dG9a signals were detected exclusively in nuclei, suggesting a regulatory role. However, mono-, di-, and trimethylated H3K9 signals were not extensively decreased in a homozygous dG9a null mutant throughout these stages. In contrast, mono- and trimethylated H3K9 signals were extensively decreased in a heterozygous DmSetdb1 mutant during spermatogenesis, and similar reduction in monomethylated H3K9 signals was observed in a homozygous Su(var)3–9 mutant. Therefore, DmSETDB1 is likely to be mainly responsible for mono- and trimethylation of H3K9 and SU(VAR)3–9 for monomethylation of H3K9 during spermatogenesis. However, the reduced methylation of H3K9 in premeiotic spermatocytes did not influence X–Y chromosome disjunction in male meiosis, suggesting that it may not be critical for spermatogenesis in Drosophila.

Keywords

histone H3K9 histone methyltransferase spermatogenesis male meiosis Drosophila 

Abbreviations

HMTase

Histone methyltransferase

H3K9

Histone H3 lysine 9

PVDF

Polyvinylidene difluoride

DAPI

4′,6-Diamidino-2-phenylindole

SDS

Sodium dodecyl sulfate

PAGE

Polyacrylamide gel electrophoresis

IgG

Immunoglobulin G

Notes

Acknowledgments

We are grateful to Drs. P. Spierer, S. Bontron, and T. Hazelrigg for fly stocks; M. Moore for English usage of the manuscript. This work was supported in part by Grants-in-Aid from JSPS, JST, and the Ministry of Education, Science, Sports, and Culture of Japan.

Supplementary material

10577_2012_9276_MOESM1_ESM.doc (32 kb)
Fig. S1 Alignment of amino acid residues of human and Drosophila histone H3 based on the CLUSTAL 2.0.12 multiple sequence alignment. The N-terminal region corresponding to the synthetic peptide used for production of monoclonal antibodies is underlined. The histone H3 lysine 9 residue is marked (DOC 32 kb)
10577_2012_9276_MOESM2_ESM.doc (64 kb)
Fig. S2 Relative mRNA levels for Su(var)3–9 and DmSETDB1 in Canton S testes. Testes and carcasses were obtained from 50 adult males (Canton S), and mRNA levels were measured by quantitative RT-PCR. The mRNA levels (%) are shown relative to those in the whole adult male. Mean values with standard deviation are shown (DOC 64 kb)

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Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Yuta Ushijima
    • 1
  • Yoshihiro H. Inoue
    • 2
  • Takahiro Konishi
    • 1
  • Daishi Kitazawa
    • 2
  • Hideki Yoshida
    • 1
    • 2
  • Kouhei Shimaji
    • 1
  • Hiroshi Kimura
    • 3
  • Masamitsu Yamaguchi
    • 1
    • 2
    Email author
  1. 1.Department of Applied BiologyKyoto Institute of TechnologySakyo-kuJapan
  2. 2.Insect Biomedical Research CenterKyoto Institute of TechnologySakyo-kuJapan
  3. 3.Graduate School of Frontier BiosciencesOsaka UniversitySuitaJapan

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