Frederick National Laboratory for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, Maryland, USA.
Microbiology and Molecular Genetics, McGovern Medical School, Houston, Texas, USA.
J Bacteriol. 2020 Oct 8;202(21). doi: 10.1128/JB.00342-20.
Cell growth and division are coordinated, ensuring homeostasis under any given growth condition, with division occurring as cell mass doubles. The signals and controlling circuit(s) between growth and division are not well understood; however, it is known in that the essential GTPase Era, which is growth rate regulated, coordinates the two functions and may be a checkpoint regulator of both. We have isolated a mutant of Era that separates its effect on growth and division. When overproduced, the mutant protein Era647 is dominant to wild-type Era and blocks division, causing cells to filament. Multicopy suppressors that prevent the filamentation phenotype of Era647 either increase the expression of FtsZ or decrease the expression of the Era647 protein. Excess Era647 induces complete delocalization of Z rings, providing an explanation for why Era647 induces filamentation, but this effect is probably not due to direct interaction between Era647 and FtsZ. The hypermorphic * allele at the native locus can suppress the effects of Era647 overproduction, indicating that extra FtsZ is not required for the suppression, but another hypermorphic allele that accelerates cell division through periplasmic signaling, *, cannot. Together, these results suggest that Era647 blocks cell division by destabilizing the Z ring. All cells need to coordinate their growth and division, and small GTPases that are conserved throughout life play a key role in this regulation. One of these, Era, provides an essential function in the assembly of the 30S ribosomal subunit in , but its role in regulating cell division is much less well understood. Here, we characterize a novel dominant negative mutant of Era (Era647) that uncouples these two activities when overproduced; it inhibits cell division by disrupting assembly of the Z ring, without significantly affecting ribosome production. The unique properties of this mutant should help to elucidate how Era regulates cell division and coordinates this process with ribosome biogenesis.
细胞的生长和分裂是协调的,确保在任何给定的生长条件下保持内稳态,分裂发生在细胞质量加倍时。生长和分裂之间的信号和控制回路尚不清楚;然而,已知在 中,必需的 GTPase Era 受生长速率调节,协调这两个功能,并且可能是两者的检查点调节剂。我们已经分离出一种分离 Era 对生长和分裂的影响的突变体。当过度表达时,突变体蛋白 Era647 对野生型 Era 具有显性并阻断分裂,导致细胞丝状化。防止 Era647 丝状化表型的多拷贝抑制子要么增加 FtsZ 的表达,要么降低 Era647 蛋白的表达。过量的 Era647 诱导 Z 环完全去定位,这解释了为什么 Era647 诱导丝状化,但这种效应可能不是由于 Era647 与 FtsZ 之间的直接相互作用。天然基因座上的超突变 * 等位基因可以抑制 Era647 过表达的影响,表明抑制不需要额外的 FtsZ,但另一个通过周质信号加速细胞分裂的超突变等位基因 * 不能。这些结果表明,Era647 通过破坏 Z 环使细胞分裂不稳定。所有细胞都需要协调它们的生长和分裂,并且在整个生命过程中保守的小 GTPases 在这种调节中发挥关键作用。其中之一,Era,在 的 30S 核糖体亚基组装中提供了必不可少的功能,但它在调节 细胞分裂中的作用知之甚少。在这里,我们描述了 Era 的一种新型显性负突变体(Era647),当过度表达时,它可以将这两种活性解耦;它通过破坏 Z 环的组装来抑制细胞分裂,而不会显著影响核糖体的产生。这种突变体的独特特性应该有助于阐明 Era 如何调节细胞分裂,并将这个过程与核糖体生物发生协调起来。