Department of Molecular Biology, University of Wyoming, Laramie, WY, 82072, USA.
Mol Microbiol. 2019 Jun;111(6):1700-1714. doi: 10.1111/mmi.14247. Epub 2019 Apr 2.
In most rod-shaped bacteria, DNA replication is quickly followed by chromosome segregation, when one of the newly duplicated centromeres moves across the cell to the opposite (or 'new') pole. Two proteins in Caulobacter crescentus, PopZ and TipN, provide directional cues at the new pole that guide the translocating chromosome to its destination. We show that centromere translocation can be inhibited by an evolutionarily conserved pole-localized protein that we have named SpbR. When overproduced, SpbR exhibits aberrant accumulation at the old pole, where it physically interacts with PopZ. This prevents the relocation of PopZ to the new pole, thereby eliminating a positional cue for centromere translocation. Consistent with this, the centromere translocation phenotype of SpbR overproducing cells is strongly enhanced in a ∆tipN mutant background. We find that pole-localized SpbR is normally cleared by ClpXP-mediated proteolysis before the time of chromosome segregation, indicating that SpbR turnover is part of the cell cycle-dependent program of polar development. This work demonstrates the importance of proteolysis as a housekeeping activity that removes outgoing factors from the developing cell pole, and provides an example of a substrate that can inhibit polar functions if it is insufficiently cleared.
在大多数杆状细菌中,DNA 复制后很快就会进行染色体分离,此时新复制的两个着丝粒之一会穿过细胞移动到相对的(或“新”的)极点。新月柄杆菌中的两种蛋白质,PopZ 和 TipN,在新极点提供定向线索,引导正在转移的染色体到达其目的地。我们表明,着丝粒易位可以被一种进化上保守的、定位于极点的蛋白质 SpbR 抑制。当 SpbR 过表达时,它会异常地积累在旧极点上,并与 PopZ 发生物理相互作用。这阻止了 PopZ 向新极点的重新定位,从而消除了着丝粒易位的位置线索。与此一致的是,在 ∆tipN 突变体背景下,SpbR 过表达细胞的着丝粒易位表型得到了强烈增强。我们发现,在染色体分离之前,定位于极点的 SpbR 通常会被 ClpXP 介导的蛋白酶体降解清除,这表明 SpbR 的周转是细胞周期依赖性极发育程序的一部分。这项工作证明了蛋白酶体作为一种管家活性的重要性,它可以将即将离开的因子从发育中的细胞极点中清除,并且提供了一个例子,说明如果底物不能被充分清除,它可能会抑制极的功能。