Gurung Pratima, McGee James P, Dvorin Jeffrey D
Division of Infectious Diseases, Boston Children's Hospital, Boston, M.A.
Department of Pediatrics, Harvard Medical School, Boston, M.A.
bioRxiv. 2024 Feb 29:2024.02.26.582160. doi: 10.1101/2024.02.26.582160.
Condensin I is a pentameric complex that regulates the mitotic chromosome assembly in eukaryotes. The kleisin subunit CAP-H of the condensin I complex acts as a linchpin to maintain the structural integrity and loading of this complex on mitotic chromosomes. This complex is present in all eukaryotes and has recently been identified in . However, how this complex is assembled and whether the kleisin subunit is critical for this complex in these parasites is yet to be explored. To examine the role of PfCAP-H during cell division within erythrocytes, we generated an inducible PfCAP-H knockout parasite. We find that PfCAP-H is dynamically expressed during mitosis with the peak expression at the metaphase plate. PfCAP-H interacts with PfCAP-G and is a non-SMC member of the condensin I complex. Notably, the absence of PfCAP-H does not alter the expression of PfCAP-G but affects its localization at the mitotic chromosomes. While mitotic spindle assembly is intact in PfCAP-H deficient parasites, duplicated centrosomes remain clustered over the mass of unsegmented nuclei with failed karyokinesis. This failure leads to the formation of an abnormal nuclear mass, while cytokinesis occurs normally. Altogether, our data suggest that PfCAP-H plays a crucial role in maintaining the structural integrity of the condensin I complex on the mitotic chromosomes and is essential for the asexual development of malarial parasites.
凝聚素I是一种五聚体复合物,可调节真核生物中的有丝分裂染色体组装。凝聚素I复合物的kleisin亚基CAP-H作为关键成分,维持该复合物在有丝分裂染色体上的结构完整性和负载。这种复合物存在于所有真核生物中,最近在……中被鉴定出来。然而,这种复合物如何组装以及kleisin亚基在这些寄生虫中对该复合物是否至关重要,仍有待探索。为了研究PfCAP-H在红细胞内细胞分裂过程中的作用,我们构建了一种可诱导的PfCAP-H基因敲除寄生虫。我们发现PfCAP-H在有丝分裂期间动态表达,在中期板处表达达到峰值。PfCAP-H与PfCAP-G相互作用,是凝聚素I复合物的非SMC成员。值得注意的是,PfCAP-H的缺失不会改变PfCAP-G的表达,但会影响其在有丝分裂染色体上的定位。虽然PfCAP-H缺陷型寄生虫中的有丝分裂纺锤体组装完好,但复制的中心体仍聚集在未分割的细胞核团上,核分裂失败。这种失败导致形成异常的核团,而胞质分裂正常发生。总之,我们的数据表明PfCAP-H在维持有丝分裂染色体上凝聚素I复合物的结构完整性方面起着关键作用,并且对疟原虫的无性发育至关重要。