Department of Biology, Center for Cancer and Developmental Biology, California State University Northridge, Northridge, CA 91330-8303, USA.
Mental Health Research Institute, Melbourne Brain Centre, Parkville, Victoria 3052, Australia.
Microbiology (Reading). 2012 Oct;158(Pt 10):2492-2503. doi: 10.1099/mic.0.055285-0. Epub 2012 Jul 12.
The master regulator CtrA oscillates during the Caulobacter cell cycle due to temporally regulated proteolysis and transcription. It is proteolysed during the G1-S transition and reaccumulates in predivisional cells as a result of transcription from two sequentially activated promoters, P1 and P2. CtrA reinforces its own synthesis by directly mediating the activation of P2 concurrently with repression of P1. To explore the role of P1 in cell cycle control, we engineered a mutation into the native ctrA locus that prevents transcription from P1 but not P2. As expected, the ctrA P1 mutant exhibits striking growth, morphological and DNA replication defects. Unexpectedly, we found CtrA and its antagonist SciP, but not DnaA, GcrA or CcrM accumulation to be dramatically reduced in the ctrA P1 mutant. SciP levels closely paralleled CtrA accumulation, suggesting that CtrA acts as a rheostat to modulate SciP abundance. Furthermore, the reappearance of CtrA and CcrM in predivisional cells was delayed in the P1 mutant by 0.125 cell cycle unit in synchronized cultures. High levels of ccrM transcription despite low levels of CtrA and increased transcription of ctrA P2 in the ctrA P1 mutant are two examples of robustness in the cell cycle. Thus, Caulobacter can adjust regulatory pathways to partially compensate for reduced and delayed CtrA accumulation in the ctrA P1 mutant.
主调控因子 CtrA 在钙杆菌细胞周期中由于受时间调控的蛋白水解和转录而发生振荡。它在 G1-S 转换期间被蛋白水解,并且由于来自两个连续激活的启动子 P1 和 P2 的转录而在预分裂细胞中重新积累。CtrA 通过直接介导 P2 的激活同时抑制 P1 的转录来增强自身的合成。为了探索 P1 在细胞周期控制中的作用,我们对天然 ctrA 基因座进行了工程改造,使其不能从 P1 转录但可以从 P2 转录。正如预期的那样,ctrA P1 突变体表现出明显的生长、形态和 DNA 复制缺陷。出乎意料的是,我们发现 CtrA 和它的拮抗剂 SciP,但不是 DnaA、GcrA 或 CcrM 的积累在 ctrA P1 突变体中显著减少。SciP 水平与 CtrA 积累密切平行,表明 CtrA 作为变阻器来调节 SciP 的丰度。此外,在同步培养物中,P1 突变体中 CtrA 和 CcrM 在预分裂细胞中的重新出现延迟了 0.125 个细胞周期单元。尽管 CtrA 水平低且 ctrA P2 的转录增加,但 ccrM 的转录水平仍然很高,这是细胞周期中稳健性的两个例子。因此,钙杆菌可以调整调控途径,以部分补偿 ctrA P1 突变体中 CtrA 积累减少和延迟的情况。