Department of Biochemistry and Molecular Biology, The Uniformed Services University of the Health Sciences-F. Edward Hébert School of Medicine, Bethesda, MD 20814, USA.
Department of Entomology and Plant Pathology, University of Arkansas Systems Division of Agriculture, Fayetteville, AR 72701, USA.
Genetics. 2023 Dec 6;225(4). doi: 10.1093/genetics/iyad169.
Functions of protein SUMOylation remain incompletely understood in different cell types. Via forward genetics, here we identified ubaBQ247*, a loss-of-function mutation in a SUMO activation enzyme UbaB in the filamentous fungus Aspergillus nidulans. The ubaBQ247*, ΔubaB, and ΔsumO mutants all produce abnormal chromatin bridges, indicating the importance of SUMOylation in the completion of chromosome segregation. The bridges are enclosed by nuclear membrane containing peripheral nuclear pore complex proteins that normally get dispersed during mitosis, and the bridges are also surrounded by cytoplasmic microtubules typical of interphase cells. Time-lapse sequences further indicate that most bridges persist through interphase prior to the next mitosis, and anaphase chromosome segregation can produce new bridges that persist into the next interphase. When the first mitosis happens at a higher temperature of 42°C, SUMOylation deficiency produces not only chromatin bridges but also many abnormally shaped single nuclei that fail to divide. UbaB-GFP localizes to interphase nuclei just like the previously studied SumO-GFP, but the nuclear signals disappear during mitosis when the nuclear pores are partially open, and the signals reappear after mitosis. The nuclear localization is consistent with many SUMO targets being nuclear proteins. Finally, although the budding yeast SUMOylation machinery interacts with LIS1, a protein critical for dynein activation, loss of SUMOylation does not cause any obvious defect in dynein-mediated transport of nuclei and early endosomes, indicating that SUMOylation is unnecessary for dynein activation in A. nidulans.
蛋白质 SUMOylation 的功能在不同的细胞类型中仍不完全了解。通过正向遗传学,我们在这里鉴定了 ubaBQ247*,这是丝状真菌构巢曲霉中 SUMO 激活酶 UbaB 的功能丧失突变。ubaBQ247*、ΔubaB 和 ΔsumO 突变体都产生异常染色质桥,表明 SUMOylation 在完成染色体分离中的重要性。这些桥被包含周围核孔复合体蛋白的核膜包围,这些蛋白在有丝分裂期间通常会分散,桥也被典型的有丝分裂间期细胞质微管包围。延时序列进一步表明,大多数桥在进入下一次有丝分裂之前在有丝分裂间期持续存在,并且后期染色体分离可以产生持续到下一次有丝分裂的新桥。当第一次有丝分裂发生在较高的 42°C 温度下时,SUMOylation 缺乏不仅会产生染色质桥,还会产生许多异常形状的单核细胞,这些细胞无法分裂。UbaB-GFP 与之前研究的 SumO-GFP 一样定位于有丝分裂间期核,但当核孔部分打开时,核信号在有丝分裂期间消失,核分裂后信号再次出现。核定位与许多 SUMO 靶标是核蛋白一致。最后,尽管酿酒酵母 SUMOylation 机制与 LIS1 相互作用,LIS1 是一种对动力蛋白激活至关重要的蛋白质,但 SUMOylation 的缺失不会导致核和早期内体的动力蛋白介导运输出现任何明显缺陷,表明在构巢曲霉中,SUMOylation 对于动力蛋白的激活不是必需的。