Bennett Richard J, Miller Mathew G, Chua Penelope R, Maxon Mary E, Johnson Alexander D
Department of Microbiology and Immunology, UCSF, San Francisco, CA 94143, USA.
Mol Microbiol. 2005 Feb;55(4):1046-59. doi: 10.1111/j.1365-2958.2005.04466.x.
It is now well established that mating can occur between diploid a and alpha cells of Candida albicans. There is, however, controversy over when, and with what efficiency, nuclear fusion follows cell fusion to create stable tetraploid a/alpha cells. In this study, we have analysed the mating process between C. albicans strains using both cytological and genetic approaches. Using strains derived from SC5314, we used a number of techniques, including time-lapse microscopy, to demonstrate that efficient nuclear fusion occurs in the zygote before formation of the first daughter cell. Consistent with these observations, zygotes micromanipulated from mating mixes gave rise to mononuclear tetraploid cells, even when no selection for successful mating was applied to them. Mating between different clinical isolates of C. albicans revealed that while all isolates could undergo nuclear fusion, the efficiency of nuclear fusion varied in different crosses. We also show that nuclear fusion in C. albicans requires the Kar3 microtubule motor protein. Deletion of the CaKAR3 gene from both mating partners had little or no effect on zygote formation but reduced the formation of stable tetraploids more than 600-fold, as determined by quantitative mating assays. These findings demonstrate that nuclear fusion is an active process that can occur in C. albicans at high frequency to produce stable, mononucleate mating products.
目前已经明确,白色念珠菌的二倍体a细胞和α细胞之间能够发生交配。然而,对于细胞融合后何时发生核融合以及核融合以何种效率产生稳定的四倍体a/α细胞,仍存在争议。在本研究中,我们使用细胞学和遗传学方法分析了白色念珠菌菌株之间的交配过程。利用源自SC5314的菌株,我们采用了多种技术,包括延时显微镜观察,以证明在第一个子细胞形成之前,合子中就发生了高效的核融合。与这些观察结果一致,即使未对它们进行成功交配的筛选,从交配混合物中显微操作得到的合子也能产生单核四倍体细胞。对不同白色念珠菌临床分离株之间的交配研究表明,虽然所有分离株都能进行核融合,但不同杂交组合中核融合的效率有所不同。我们还表明,白色念珠菌中的核融合需要Kar3微管运动蛋白。通过定量交配试验确定,从两个交配伙伴中删除CaKAR3基因对合子形成几乎没有影响,但使稳定四倍体的形成减少了600多倍。这些发现表明,核融合是一个活跃的过程,在白色念珠菌中能够以高频率发生,从而产生稳定的单核交配产物。