Morelle Christelle, Sterkers Yvon, Crobu Lucien, MBang-Benet Diane-Ethna, Kuk Nada, Portalès Pierre, Bastien Patrick, Pagès Michel, Lachaud Laurence
Laboratory of Parasitology-Mycology, Faculty of Medicine, University Montpellier 1, Montpellier F34090, France CNRS 5290-IRD 224-University Montpellier 1&2 (UMR 'MiVEGEC'), Montpellier F34090, France Department of Parasitology-Mycology, University Hospital Centre (CHU), Montpellier F34090, France.
CNRS 5290-IRD 224-University Montpellier 1&2 (UMR 'MiVEGEC'), Montpellier F34090, France.
Nucleic Acids Res. 2015 Apr 30;43(8):4013-27. doi: 10.1093/nar/gkv056. Epub 2015 Feb 17.
Nucleoporins are evolutionary conserved proteins mainly involved in the constitution of the nuclear pores and trafficking between the nucleus and cytoplasm, but are also increasingly viewed as main actors in chromatin dynamics and intra-nuclear mitotic events. Here, we determined the cellular localization of the nucleoporin Mlp2 in the 'divergent' eukaryotes Leishmania major and Trypanosoma brucei. In both protozoa, Mlp2 displayed an atypical localization for a nucleoporin, essentially intranuclear, and preferentially in the periphery of the nucleolus during interphase; moreover, it relocated at the mitotic spindle poles during mitosis. In T. brucei, where most centromeres have been identified, TbMlp2 was found adjacent to the centromeric sequences, as well as to a recently described unconventional kinetochore protein, in the periphery of the nucleolus, during interphase and from the end of anaphase onwards. TbMlp2 and the centromeres/kinetochores exhibited a differential migration towards the poles during mitosis. RNAi knockdown of TbMlp2 disrupted the mitotic distribution of chromosomes, leading to a surprisingly well-tolerated aneuploidy. In addition, diploidy was restored in a complementation assay where LmMlp2, the orthologue of TbMlp2 in Leishmania, was expressed in TbMlp2-RNAi-knockdown parasites. Taken together, our results demonstrate that Mlp2 is involved in the distribution of chromosomes during mitosis in trypanosomatids.
核孔蛋白是进化上保守的蛋白质,主要参与核孔的构成以及细胞核与细胞质之间的物质运输,但也越来越被视为染色质动态变化和核内有丝分裂事件的主要参与者。在这里,我们确定了核孔蛋白Mlp2在“分化”真核生物利什曼原虫和布氏锥虫中的细胞定位。在这两种原生动物中,Mlp2表现出核孔蛋白的非典型定位,主要位于核内,在间期优先位于核仁周边;此外,在有丝分裂期间它重新定位到有丝分裂纺锤体极。在已鉴定出大多数着丝粒的布氏锥虫中,在间期以及从后期结束开始,发现TbMlp2在核仁周边与着丝粒序列相邻,以及与最近描述的一种非常规动粒蛋白相邻。在有丝分裂期间,TbMlp2和着丝粒/动粒向两极表现出不同的迁移。RNA干扰敲低TbMlp2会破坏染色体的有丝分裂分布,导致令人惊讶的耐受性良好的非整倍体。此外在互补试验中恢复了二倍体状态,在该试验中,布氏锥虫TbMlp2的直系同源物利什曼原虫LmMlp2在TbMlp2-RNA干扰敲低的寄生虫中表达。综上所述,我们的结果表明Mlp2参与了锥虫有丝分裂期间染色体的分布。