Characterization of fission yeast meiotic mutants based on live observation of meiotic prophase nuclear movement
Characterization of fission yeast meiotic mutants based on live observation of meiotic prophase nuclear movement
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DOI：
10.1007/s004120050417
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发表时间：
2000-04-01
期刊：
影响因子：
1.6
通讯作者：
Chikashige, Y
中科院分区：
文献类型：
--
作者：
Hiraoka, Y;Ding, DQ;Chikashige, Y
通过对裂殖酵母裂殖酵母细胞核运动的实时观察，对裂殖酵母裂殖酵母的四个减数分裂突变体进行了鉴定。用DNA特异性荧光染料Hoechst 33342或与DNA聚合酶α的N-末端部分融合的水母绿色荧光蛋白（GFP）对细胞核进行染色。我们首先跟踪野生型细胞的核动力学，以确定减数分裂事件的时间顺序：结合合子中的核融合立即发生振荡核运动，持续146分钟;然后，在休息后，核在第一次减数分裂前在细胞中心停留26分钟。接下来，我们研究了四个减数分裂突变体的核动力学：mei 1（也称为mat 2），mei 4，dhc 1和taz 1。Mei 1和Mei 4都在减数分裂前期被捕，然而，我们的观察表明，Mei 1逮捕的时间是完全不同的Mei 4：Mei 1突变逮捕核融合后，但开始振荡核运动之前，而Mei 4突变逮捕后，核已经完成了振荡运动，但在第一次减数分裂。我们还展示了dhc 1和taz 1的动态表型的例子，它们都完成了减数分裂，但表现出受损的核运动和同源重组的频率降低：dhc 1突变体表现出核融合后没有核运动，而taz 1突变体表现出严重受损的核融合后的核运动。
We characterized four meiotic mutants of the fission yeast Schizosaccharomyces pombe by live observation of nuclear movement. Nuclei were stained with either the DNA-specific fluorescent dye Hoechst 33342 or jellyfish green fluorescent protein (GFP) fused with the N-terminal portion of DNA polymerase alpha. We first followed nuclear dynamics in wild-type cells to determine the temporal sequence of meiotic events: nuclear fusion in the conjugated zygote is immediately followed by oscillatory nuclear movements that continue for 146 min; then, after coming to rest, the nucleus remains in the center of the cell for 26 min before the first meiotic division. Next we examined nuclear dynamics in four meiotic mutants: mei1 (also called mat2), mei4, dhc1, and taz1. Mei1 and mei4 both arrest during meiotic prophase; our observations, however, show that the timing of mei1 arrest is quite different from that of mei4: the mei1 mutant arrests after nuclear fusion but before starting the oscillatory nuclear movements, while the mei4 mutant arrests after the nucleus has completed the oscillatory movements but before the first meiotic division. We also show examples of the dynamic phenotypes of dhc1 and taz1, both of which complete meiosis but exhibit impaired nuclear movement and reduced frequencies of homologous recombination: the dhc1 mutant exhibits no nuclear movement after nuclear fusion, while the taz1 mutant exhibits severely impaired nuclear movement after nuclear fusion.