Center for Tropical and Emerging Global Diseases and Department of Cellular Biology, University of Georgia, Athens, GA 30602, USA.
Proc Natl Acad Sci U S A. 2011 Mar 1;108(9):3767-72. doi: 10.1073/pnas.1006741108. Epub 2011 Feb 14.
Members of the eukaryotic phylum Apicomplexa are the cause of important human diseases including malaria, toxoplasmosis, and cryptosporidiosis. These obligate intracellular parasites produce new invasive stages through a complex budding process. The budding cycle is remarkably flexible and can produce varied numbers of progeny to adapt to different host-cell niches. How this complex process is coordinated remains poorly understood. Using Toxoplasma gondii as a genetic model, we show that a key element to this coordination is the centrocone, a unique elaboration of the nuclear envelope that houses the mitotic spindle. Exploiting transgenic parasite lines expressing epitope-tagged centromeric H3 variant CenH3, we identify the centromeres of T. gondii chromosomes by hybridization of chromatin immunoprecipitations to genome-wide microarrays (ChIP-chip). We demonstrate that centromere attachment to the centrocone persists throughout the parasite cell cycle and that centromeres localize to a single apical region within the nucleus. Centromere sequestration provides a mechanism for the organization of the Toxoplasma nucleus and the maintenance of genome integrity.
真核生物门顶复门的成员是疟疾、弓形体病和隐孢子虫病等重要人类疾病的病原体。这些专性细胞内寄生虫通过复杂的出芽过程产生新的侵袭阶段。出芽周期非常灵活,可以产生不同数量的后代,以适应不同的宿主细胞小生境。这个复杂的过程是如何协调的仍然知之甚少。我们使用刚地弓形虫作为遗传模型,表明这种协调的一个关键因素是中心体,这是核膜的一个独特结构,其中包含有丝分裂纺锤体。利用表达表位标记着丝粒 H3 变体 CenH3 的转基因寄生虫系,我们通过将染色质免疫沉淀与全基因组微阵列(ChIP-chip)杂交来鉴定 T. gondii 染色体的着丝粒。我们证明着丝粒附着在中心体上贯穿整个寄生虫细胞周期,并且着丝粒定位于核内的单个顶端区域。着丝粒的隔离为刚地弓形虫核的组织和基因组完整性的维持提供了一种机制。