Liu Mengying, Wang Li, Chen Wenli
State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China.
Wei Sheng Wu Xue Bao. 2010 Jan;50(1):36-40.
NtcA is a master transcriptional regulator in global nitrogen control in Anabaena. Heterocyst differentiation depends on the NtcA protein. A large number of genes regulated by NtcA have a consensus sequence, GTA-N8-TAC, for NtcA binding. Bioinformatics analysis of alr1390 in the chromosome of Anabaena sp. PCC 7120 indicates that there is a conservative NtcA binding box, GTA-TAGTTTTC-TAC, upstream the putative tsp position of alr1390.
To understand the relationship between alr1390 and NtcA, METHODS: Real-time RT-PCR and EMSA were used.
Real-time RT-PCR results showed that the transcriptional level of alr1390 remained constant before and after combined nitrogen step-down in wild type Anabaena sp. PCC 7120. However, in the ntcA mutant, there was an increase in the transcriptional level of alr1390 after 12h of combined nitrogen step-down, which indicated that alr1390 should be subject to the regulation of NtcA. Additionally although no conspicuous NtcA-alr1390 complex was detected by EMSA in vitro, a smear appeared in gel electrophoresis.
These results suggested that alr1390 might be regulated by NtcA.
NtcA是鱼腥藻全局氮控制中的主要转录调节因子。异形胞分化依赖于NtcA蛋白。大量受NtcA调控的基因具有NtcA结合的共有序列GTA-N8-TAC。对鱼腥藻PCC 7120染色体上alr1390的生物信息学分析表明,在alr1390假定的转录起始点上游存在一个保守的NtcA结合框GTA-TAGTTTTC-TAC。
了解alr1390与NtcA之间的关系。
采用实时RT-PCR和电泳迁移率变动分析(EMSA)。
实时RT-PCR结果显示,在野生型鱼腥藻PCC 7120中,联合氮素浓度降低前后,alr1390的转录水平保持恒定。然而,在ntcA突变体中,联合氮素浓度降低12小时后,alr1390的转录水平有所增加,这表明alr1390应该受NtcA的调控。此外,尽管在体外EMSA中未检测到明显的NtcA-alr1390复合物,但在凝胶电泳中出现了拖尾现象。
这些结果表明alr1390可能受NtcA调控。