Lin Gui-Ming, Zhang Ju-Yuan, Shao Zhi-Hui, Yang Chen, Zhao Guo-Ping, Huang Kai-Yao, Zhang Cheng-Cai
State Key Laboratory of Freshwater Ecology and Biotechnology, Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430070, Hubei, China.
State Key Laboratory of Freshwater Ecology and Biotechnology, Key Laboratory of Algal Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430070, Hubei, China.
Microbiol Res. 2025 Jan;290:127970. doi: 10.1016/j.micres.2024.127970. Epub 2024 Nov 9.
PacR (All3953) has previously been identified as a global transcriptional regulator of carbon assimilation in cyanobacteria. In the facultative diazotrophic and filamentous cyanobacterium Anabaena PCC 7120 (Anabaena), inactivation of pacR has been shown to affect cell growth under various conditions. Nitrogen fixation in Anabaena occurs in heterocysts, cells differentiated semiregularly along the filaments following deprivation of combined nitrogen such as nitrate or ammonium. Here, we created a markerless deletion mutant of pacR. In addition to its growth defects observed under different light and nitrogen conditions, the mutant could form a high frequency of heterocysts, including heterocyst doublets, even in the presence of nitrate. Inactivation of pacR led to the upregulation of ntcA, a global regulator of nitrogen metabolism and heterocyst formation, as well as downregulation of genes involved in nitrate uptake and assimilation. These changes led to N-limited cells in the presence of nitrate. PacR also regulates most of the genes encoding bicarbonate transport systems. The promoter regions of ntcA, and several other genes involved in nitrogen or carbon uptake and assimilation, as well as patS and hetN involved in heterocyst patterning can be directly recognized by PacR in vitro. These findings, along with previously reported ChIP-seq data, establish PacR as a crucial transcriptional regulator for balancing carbon and nitrogen metabolism in cyanobacteria.
PacR(All3953)先前已被鉴定为蓝藻中碳同化的全局转录调节因子。在兼性固氮丝状蓝藻鱼腥藻PCC 7120(鱼腥藻)中,pacR的失活已被证明会在各种条件下影响细胞生长。鱼腥藻中的固氮作用发生在异形胞中,异形胞是在剥夺硝酸盐或铵等化合态氮后沿丝状体半规则分化的细胞。在此,我们创建了pacR的无标记缺失突变体。除了在不同光照和氮条件下观察到的生长缺陷外,该突变体即使在存在硝酸盐的情况下也能形成高频率的异形胞,包括异形胞双联体。pacR的失活导致ntcA上调,ntcA是氮代谢和异形胞形成的全局调节因子,同时也导致参与硝酸盐吸收和同化的基因下调。这些变化导致在存在硝酸盐的情况下细胞处于氮限制状态。PacR还调节大多数编码碳酸氢盐转运系统的基因。ntcA的启动子区域,以及其他一些参与氮或碳吸收和同化的基因,以及参与异形胞模式形成的patS和hetN,在体外可被PacR直接识别。这些发现,连同先前报道的ChIP-seq数据,确立了PacR作为蓝藻中平衡碳和氮代谢的关键转录调节因子的地位。