Identification of Nuclear Localization Signals of Drosophila G9a Histone H3 Methyltransferase

Kato, Yasuko; Ushijima, Yuta; Yamaguchi, Masamitsu

doi:10.1247/csf.10027

Cited by 3 publications

(5 citation statements)

References 21 publications

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“…Across these cycles, dG9a could be responsible for regulating the expression level of many genes required for embryogenesis and transcription. Human G9a localizes in nuclei of HEK293 cells (Tachibana et al 2001) and is also reported to be present in nuclei in salivary glands (Stabell et al 2006;Kato et al 2008) and in cultured Drosophila Kc cells (Kato et al 2011). However, as found here, dG9a almost exclusively localizes in cytoplasm of early embryos up to nuclear division cycle 7 (Fig.…”

Section: Discussionsupporting

confidence: 77%

“…Nuclear localization signals that are distinct from those of mammalian G9a in their positions and amino acid sequences may be responsible for the nuclear localization of dG9a during cycle 8 (Kato et al . ). It is well known that a first wave of ZGA occurs around cycle 8 and activates genes required for cellularization (De Renzis et al .…”

Section: Discussionmentioning

confidence: 97%

“…) and in cultured Drosophila Kc cells (Kato et al . ). However, as found here, dG9a almost exclusively localizes in cytoplasm of early embryos up to nuclear division cycle 7 (Fig.…”

Section: Discussionmentioning

confidence: 97%

“…In later stages, dG9a signals rather accumulate in nuclei from nuclear division cycles 8 to 12 and appear to be responsible for di-methylation of nuclear H3K9. Nuclear localization signals that are distinct from those of mammalian G9a in their positions and amino acid sequences may be responsible for the nuclear localization of dG9a during cycle 8 (Kato et al 2011). It is well known that a first wave of ZGA occurs around cycle 8 and activates genes required for cellularization (De Renzis et al 2007).…”

Section: Discussionmentioning

confidence: 99%

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Genomewide identification of target genes of histone methyltransferase dG9a during Drosophila embryogenesis

et al. 2015

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Post-translational modification of the histone plays important roles in epigenetic regulation of various biological processes. Among the identified histone methyltransferases (HMTases), G9a is a histone H3 Lys 9 (H3K9)-specific example active in euchromatic regions. Drosophila G9a (dG9a) has been reported to feature H3K9 dimethylation activity in vivo. Here, we show that the time required for hatching of a homozygous dG9a null mutant and heteroallelic combination of dG9a null mutants is delayed, suggesting that dG9a is at least partially responsible for progression of embryogenesis. Immunocytochemical analyses of the wild-type and the dG9a null mutant flies indicated that dG9a localizes in cytoplasm up to nuclear division cycle 7 where it is likely responsible for di-methylation of nucleosome-free H3K9. From cycles 8-11, dG9a moves into the nucleus and is responsible for di-methylating H3K9 in nucleosomes. RNA-sequence analysis utilizing early wild-type and dG9a mutant embryos showed that dG9a down-regulates expression of genes responsible for embryogenesis. RNA fluorescent in situ hybridization analysis further showed temporal and spatial expression patterns of these mRNAs did not significantly change in the dG9a mutant. These results indicate that dG9a controls transcription levels of some zygotic genes without changing temporal and spatial expression patterns of the transcripts of these genes.

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Section: Discussionsupporting

confidence: 77%

Section: Discussionmentioning

confidence: 97%

“…) and in cultured Drosophila Kc cells (Kato et al . ). However, as found here, dG9a almost exclusively localizes in cytoplasm of early embryos up to nuclear division cycle 7 (Fig.…”

Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

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Genomewide identification of target genes of histone methyltransferase dG9a during Drosophila embryogenesis

et al. 2015

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“…We prefer to support the first speculation, not only because the mouse indeed contained two NLS but also because these signals were conserved in mammals [15]. Even in Drosophila, the nuclear localization signal of G9a has been identified [34], which suggests that the cell nucleus localization of G9a is highly conserved and that its own biological function is very important. In addition, in evolutionary terms, an organism would not produce a protein and then degrade it repeatedly within the limited time frame because it would be an extreme waste of biosynthetic energy [35].…”

Section: Discussionmentioning

confidence: 94%

G9a co-localized with histone H3 lysine 9 monomethylation but not dimethylation in a nuclear membrane-dependent manner during mouse preimplantation embryo development

Tang²,

Chen

et al. 2012

J Assist Reprod Genet

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RecQ5 Protein Translocation into the Nucleus by a Nuclear Localization Signal

Sakurai

Tsutsui

Higashi

et al. 2013

Biological & Pharmaceutical Bulletin

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RecQ5, a member of the RecQ helicase family, maintains genome stability via participation in many DNA metabolic processes including DNA repair, DNA resolution, and RNA transcription, processes occurring in the nucleus. Previously, we reported that RecQ5 and Rad51, also involved in DNA repair, become colocalized in nuclei when co-expressed in cultured cells. Nuclear localization of RecQ5 appears to be important for cellular function along with Rad51. However, little is known about the nuclear localization of RecQ5. Here, we generated enhanced green fluorescent protein (EGFP)-tagged RecQ5 transgenic flies and analyzed localization of this protein in early embryos by live imaging. In syncytial embryos, RecQ5 was localized synchronously in interphase nuclei, and spread repeatedly over the embryos in mitosis. Thus, RecQ5 was transported into nuclei at the early interphase. Furthermore, we examined the subcellular localization of a series of truncated forms of Drosophila RecQ5 in cultured cells to determine the nuclear localization signal (NLS). Entire coding or deleted RecQ5 sequences of various sizes were ligated into EGFP vectors, which were then used to transfect cultured Drosophila cells. The region responsible for nuclear localization of Drosophila RecQ5 contained a short stretch of positively charged basic amino acids, 2 of which were particularly important for the nuclear localization. This stretch was sufficient for nuclear localization when fused with EGFP. Although the NLS of Drosophila RecQ5 was distinct from that of human RECQL5 in terms of position and amino acid sequence, this fly RecQ5 protein was translocated into the nucleus by an NLS. Key words RecQ; nuclear localization signal; DrosophilaRecQ5 DNA helicase is a member of the RecQ family, in which there are 5 members in humans. Three of them are involved in predisposition to cancer, premature aging and/ or developmental abnormality diseases such as Bloom and Werner syndromes, which are caused by mutations in the BLM and WRN genes, respectively; and 3 other syndromes, i.e., Rothmund-Thomson, RAPADALINO, and BallerGerold syndromes, are caused by mutations in the RecQ4 gene.1-4) Although RecQL1 and RECQL5 (human homolog of Drosophila RecQ5), the other 2 members of the RecQ family in humans, have not been associated with any human genetic diseases, Recql5-knock out mice show susceptibility to cancer, 5) suggesting that RecQ5 may also play an important role in preventing cancer. Furthermore, it is postulated that RECQL5 has multiple activities in transcription with RNA pol II, 6,7) homologous recombination with MRN and Rad51, 8,9) and DNA decatenation with Topo II. 10) These activities are elicited in the nucleus. Nuclear localization of RecQ5 may be important for these activities. However, the precise nuclear transport mechanism of RecQ5 remains unknown.In Drosophila, the frequencies of spontaneous and induced chromosomal aberrations are increased in RecQ5-mutant neuroblasts.11) These data imply that double-stranded break (DSB) damage to DNA accumulates...

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Identification of Nuclear Localization Signals of Drosophila G9a Histone H3 Methyltransferase

Cited by 3 publications

References 21 publications

Genomewide identification of target genes of histone methyltransferase dG9a during Drosophila embryogenesis

Genomewide identification of target genes of histone methyltransferase dG9a during Drosophila embryogenesis

G9a co-localized with histone H3 lysine 9 monomethylation but not dimethylation in a nuclear membrane-dependent manner during mouse preimplantation embryo development

RecQ5 Protein Translocation into the Nucleus by a Nuclear Localization Signal

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