M. Goto, Narumi Toda, Kouhei Shimaji, Dang Ngoc Anh Suong, Nicole Vo, H. Kimura, H. Yoshida, Y. Inoue, M. Yamaguchi
{"title":"Polycomb-dependent nucleolus localization of Jumonji/Jarid2 during Drosophila spermatogenesis","authors":"M. Goto, Narumi Toda, Kouhei Shimaji, Dang Ngoc Anh Suong, Nicole Vo, H. Kimura, H. Yoshida, Y. Inoue, M. Yamaguchi","doi":"10.1080/21565562.2016.1232023","DOIUrl":null,"url":null,"abstract":"ABSTRACT Drosophila Jumonji/Jarid2 (dJmj) has been identified as a component of Polycomb repressive complex 2. However, it is suggested that dJmj has both PRC-dependent and –independent roles. Subcellular localization of dJmj during spermatogenesis is unknown. We therefore performed immunocytochemical analyses with specific antibodies to dJmj and tri-methylation at lysine 27 on histone H3 (H3K27me3). Interestingly, dJmj exclusively localizes at nucleolus in the late growth stage. Examination of the dJmj localization in various Polycomb group (PcG) mutant lines at the late growth stage allowed identification of some PcG genes, including Polycomb (Pc), to be responsible for dJmj recruitment to nucleolus. In addition, we found that size of nucleolus was decreased in some of these mutant lines. In a mutant of testis-specific TAF homolog (tTAF) that is responsible for nucleolus localization of Pc, dJmj signals were detected not only at nucleolus but also on the condensed chromatin in the late growth stage. Duolink In situ Proximity ligation assay clarified that Pc interacts with dJmj at nucleolus in the late growth stage. Furthermore, the level of H3K27me3 decreased in nuclei at this stage. Taken together, we conclude that tTAF is responsible for recruitments of dJmj to nucleolus in the late growth stage that appears to be mediated by Pc. Compartmentalization of dJmj in nucleolus together with some of PcG may be necessary to de-repress the expression of genes required to cellular growth and proliferation in the following meiotic divisions.","PeriodicalId":22074,"journal":{"name":"Spermatogenesis","volume":"34 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2016-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"4","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Spermatogenesis","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1080/21565562.2016.1232023","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 4
Abstract
ABSTRACT Drosophila Jumonji/Jarid2 (dJmj) has been identified as a component of Polycomb repressive complex 2. However, it is suggested that dJmj has both PRC-dependent and –independent roles. Subcellular localization of dJmj during spermatogenesis is unknown. We therefore performed immunocytochemical analyses with specific antibodies to dJmj and tri-methylation at lysine 27 on histone H3 (H3K27me3). Interestingly, dJmj exclusively localizes at nucleolus in the late growth stage. Examination of the dJmj localization in various Polycomb group (PcG) mutant lines at the late growth stage allowed identification of some PcG genes, including Polycomb (Pc), to be responsible for dJmj recruitment to nucleolus. In addition, we found that size of nucleolus was decreased in some of these mutant lines. In a mutant of testis-specific TAF homolog (tTAF) that is responsible for nucleolus localization of Pc, dJmj signals were detected not only at nucleolus but also on the condensed chromatin in the late growth stage. Duolink In situ Proximity ligation assay clarified that Pc interacts with dJmj at nucleolus in the late growth stage. Furthermore, the level of H3K27me3 decreased in nuclei at this stage. Taken together, we conclude that tTAF is responsible for recruitments of dJmj to nucleolus in the late growth stage that appears to be mediated by Pc. Compartmentalization of dJmj in nucleolus together with some of PcG may be necessary to de-repress the expression of genes required to cellular growth and proliferation in the following meiotic divisions.
果蝇Jumonji/Jarid2 (dJmj)是Polycomb抑制复合体2的一个组成部分。然而,有人认为dJmj同时具有prc依赖和不依赖的作用。精子发生过程中dJmj的亚细胞定位尚不清楚。因此,我们使用dJmj特异性抗体和组蛋白H3上赖氨酸27的三甲基化(H3K27me3)进行了免疫细胞化学分析。有趣的是,在生长后期,dJmj只局限于核仁。对生长后期不同Polycomb group (PcG)突变系中dJmj的定位进行了检测,从而确定了包括Polycomb (Pc)在内的一些PcG基因负责dJmj向核核募集。此外,我们发现一些突变系的核仁大小减小了。在负责Pc核仁定位的睾丸特异性TAF同源物(tTAF)突变体中,不仅在核仁上检测到dJmj信号,而且在生长后期的浓缩染色质上也检测到dJmj信号。Duolink原位接近连接实验表明,Pc在生长后期核仁上与dJmj相互作用。此外,细胞核中H3K27me3的水平在这一阶段下降。综上所述,我们得出结论,tTAF负责在生长后期向核仁招募dJmj,这似乎是由Pc介导的。核仁中dJmj和部分PcG的区隔化可能是抑制减数分裂后细胞生长和增殖所需基因表达的必要条件。