Sahasrabuddhe Amogh A, Nayak Ramesh C, Gupta Chhitar M
Division of Molecular and Structural Biology, Central Drug Research Institute, M.G. Marg, Lucknow 226001, India.
J Cell Sci. 2009 May 15;122(Pt 10):1691-9. doi: 10.1242/jcs.044651. Epub 2009 Apr 28.
In general, coronins play an important role in actin-based processes, and are expressed in a variety of eukaryotic cells, including Leishmania. Here, we show that Leishmania coronin preferentially distributes to the distal tip during cytokinesis, and interacts with microtubules through a microtubule-based motor, kinesin K39. We further show that reduction in coronin levels by 40-50% in heterozygous coronin mutants results in generation of bipolar cells (25-30%), specifically in the log phase, owing to unregulated growth of the corset microtubules. Further analysis of bipolar cells revealed that the main cause of generation of bipolar cell morphology is the intrusion of the persistently growing corset microtubules into the other daughter cell corset from the opposite direction. This defect in cytokinesis, however, disappears upon episomal gene complementation. Additionally, our attempts to prepare homozygous mutants were unsuccessful, as only the aneuploid cells survive the selection process. These results indicate that coronin regulates microtubule remodeling during Leishmania cytokinesis and is essentially required for survival of these parasites in culture.
一般来说,冠蛋白在基于肌动蛋白的过程中发挥重要作用,并在包括利什曼原虫在内的多种真核细胞中表达。在这里,我们表明利什曼原虫冠蛋白在胞质分裂期间优先分布于远端尖端,并通过基于微管的马达蛋白驱动蛋白K39与微管相互作用。我们进一步表明,杂合冠蛋白突变体中冠蛋白水平降低40 - 50%会导致双极细胞的产生(25 - 30%),特别是在对数期,这是由于束腰微管的无节制生长所致。对双极细胞的进一步分析表明,双极细胞形态产生的主要原因是持续生长的束腰微管从相反方向侵入另一个子细胞的束腰。然而,这种胞质分裂缺陷在附加型基因互补后消失。此外,我们制备纯合突变体的尝试未成功,因为只有非整倍体细胞在选择过程中存活下来。这些结果表明,冠蛋白在利什曼原虫胞质分裂过程中调节微管重塑,并且是这些寄生虫在培养中存活所必需的。