Genes & Genetic Systems
Online ISSN : 1880-5779
Print ISSN : 1341-7568
ISSN-L : 1341-7568
最新号
選択された号の論文の8件中1~8を表示しています
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Full papers
  • Kotaro Tsukada, Ryouhei Yoshihara, Shin Hatakeyama, Akihiko Ichiishi, ...
    原稿種別: Full paper
    2020 年 95 巻 6 号 p. 281-289
    発行日: 2020/12/01
    公開日: 2021/03/23
    [早期公開] 公開日: 2021/02/06
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    Photoreactivation is a mechanism in which photolyase directly repairs either cyclobutane pyrimidine dimers (CPDs) or (6-4) photoproducts [(6-4) PPs] caused by ultraviolet (UV) light. In the filamentous fungus Neurospora crassa, some UV-sensitive mutants such as mus-44 have been reported to exhibit a partial photoreactivation defect (PPD) phenotype, but its mechanism has not been elucidated for a long time. In this study, the N. crassa CPD photolyase PHR was overexpressed in the Δmus-44 strain, but photoreactivation ability was not increased. Furthermore, Escherichia coli CPD photolyase or Arabidopsis thaliana (6-4) PP photolyase was also introduced into Δmus-44; however, the PPD phenotype was not complemented. These results suggested that the PPD phenotype in N. crassa is not caused by residual unrepaired pyrimidine dimers, which are the main type of DNA damage caused by UV irradiation. Finally, we revealed that Δmus-44, but not the Δmus-43 strain, which does not show the PPD phenotype, displayed higher sensitivity with increasing dose rate of UV. Moreover, Δmus-44 was also sensitive to an interstrand crosslinking agent. This indicates that the high dose of UV in our experimental condition induces DNA damage other than pyrimidine dimers, and that such damage is a likely cause of the PPD phenotype.

  • Kazushi Yasuda, Yuri Kato, Shogo Ikeda, Shinji Kawano
    原稿種別: Full paper
    2020 年 95 巻 6 号 p. 291-302
    発行日: 2020/12/01
    公開日: 2021/03/23
    [早期公開] 公開日: 2021/02/06
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    Type II DNA topoisomerase (topo II) catalyzes double-stranded DNA cleavage and re-ligation, thus solving problems in DNA topology. Vertebrates have two isozymes (α and β). Recently, the C-terminal regulatory domain (CRD), which regulates catalytic activity and subnuclear localization by associating with RNA, was identified within the C-terminal domain (CTD) of rat topo IIβ. In contrast, it is unclear whether a β CRD-like domain is present in the CTD of topo IIα. In this study, we aimed to identify an RNA-mediated regulatory domain in the rat topo IIα CTD. First, we exchanged the CTDs of rat topo IIα (amino acids 1,192–1,528) and β (1,201–1,614) and examined the two chimeras’ in vitro catalytic activities. Interestingly, the relaxation activities of topo IIα WT enzyme and both of the CTD-swapped mutants were inhibited in the presence of isolated cellular RNA, suggesting that the α CTD is involved in the RNA-mediated regulation of catalytic activity in topo IIα. The results of on-bead assays using a CTD-deleted mutant of rat topo IIα indicated that the RNA-mediated inhibition of the relaxation activity was caused by an interaction between the α CTD and RNA. Further, to identify the domain within the CTD that is associated with subnuclear localization of rat topo IIα, we transiently expressed EGFP-tagged CTD deletion mutants in human cells. The data indicated that the 1,192–1,289 region of rat topo IIα was required for targeting the enzyme to nucleoli. Finally, a relaxation assay using 1–1,289 and Δ1,192–1,289 truncated mutants indicated that the 1,192–1,289 region is involved in RNA-mediated inhibition. These results indicated that the CTD of rat topo IIα, containing the 1,192–1,289 region, is involved in the regulation of catalytic activity by associating with RNA, as well as in the localization to nucleoli in interphase cells.

  • Maiko Saito, Minoru Nakayama, Kyota Fujita, Atsuko Uchida, Hiroyuki Ya ...
    原稿種別: Full paper
    2020 年 95 巻 6 号 p. 303-314
    発行日: 2020/12/01
    公開日: 2021/03/23
    [早期公開] 公開日: 2021/02/13
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    yata mutants of Drosophila melanogaster exhibit phenotypes including progressive brain shrinkage, developmental abnormalities and shortened lifespan, whereas in mammals, null mutations of the yata ortholog Scyl1 result in motor neuron degeneration. yata mutation also causes defects in the anterograde intracellular trafficking of a subset of proteins including APPL, which is the Drosophila ortholog of mammalian APP, a causative molecule in Alzheimer’s disease. SCYL1 binds and regulates the function of coat protein complex I (COPI) in secretory vesicles. Here, we reveal a role for the Drosophila YATA protein in the proper localization of COPI. Immunohistochemical analyses performed using confocal microscopy and structured illumination microscopy showed that YATA colocalizes with COPI and GM130, a cis-Golgi marker. Analyses using transgenically expressed YATA with a modified N-terminal sequence revealed that the N-terminal portion of YATA is required for the proper subcellular localization of YATA. Analysis using transgenically expressed YATA proteins in which the C-terminal sequence was modified revealed a function for the C-terminal portion of YATA in the subcellular localization of COPI. Notably, when YATA was mislocalized, it also caused the mislocalization of COPI, indicating that YATA plays a role in directing COPI to the proper subcellular site. Moreover, when both YATA and COPI were mislocalized, the staining pattern of GM130 revealed Golgi with abnormal elongated shapes. Thus, our in vivo data indicate that YATA plays a role in the proper subcellular localization of COPI.

  • Yan Luo, Jing Wang, Wei Lu, Yang Liu, Yun Huang, Dichun Luo
    原稿種別: Full paper
    2020 年 95 巻 6 号 p. 315-321
    発行日: 2020/12/01
    公開日: 2021/03/23
    [早期公開] 公開日: 2021/02/28
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    Asthma is one of the most common diseases, with a high prevalence among children. To date, systemic co-expression analysis for this disease has not been undertaken to explain its pathogenesis. Here we identified differentially expressed genes (DEGs) in 87 samples, and then constructed co-expression modules via weighted gene co-expression network analysis (WGCNA) and investigated the functional enrichment of co-expressed genes in terms of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG). Meanwhile, protein–protein interaction (PPI) network and miRNA–transcription factor–target (miRNA–TF–target) regulatory network analyses were performed to screen hub genes. As a result, 3,469 DEGs were identified in this study, of which 1,860 genes were up-regulated and 1,609 genes were down-regulated. Using WGCNA, we identified two key modules, named MEbrown and MEblue, that may play important roles in asthma. Functional enrichment analysis revealed that MEbrown was enriched in 37 KEGG pathways and 472 biological processes (BPs), while MEblue was enriched in 16 KEGG pathways and 449 BPs. From PPI and miRNA–TF–target regulatory network analysis, a total of 31 TFs, seven miRNAs and 28 nodes were identified. Our findings should provide a framework of therapeutic targets for treating children with acute asthma.

Short communication
  • Tomoya Suzuki, Akira S. Hirao, Masaki Takenaka, Koki Yano, Koji Tojo
    原稿種別: Short communication
    2020 年 95 巻 6 号 p. 323-329
    発行日: 2020/12/01
    公開日: 2021/03/23
    [早期公開] 公開日: 2021/01/24
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    We developed microsatellite markers for Appasus japonicus (Heteroptera: Belostomatidae). This belostomatid bug is distributed in East Asia (Japanese Archipelago, Korean Peninsula and mainland China) and often listed as an endangered species in the Red List or the Red Data Book at the national and local level in Japan. Here, we describe twenty novel polymorphic microsatellite loci developed for A. japonicus, and marker suitability was evaluated using 56 individuals from four A. japonicus populations (Nagano, Hiroshima and Yamaguchi prefectures, Japan, and Chungcheongnam-do, Korea). The number of alleles per locus ranged from 1 to 12 (mean = 2.5), and the average observed and expected heterozygosity and fixation index per locus were 0.270, 0.323 and 0.153, respectively. In addition, a population structure analysis was conducted using the software STRUCTURE, and its result suggested that the 20 markers described here will be useful for investigating the genetic structure of A. japonicus populations, which should contribute to population genetics studies of this species.

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