Disruption of the nitrogen regulatory gene AcareA in Acremonium chrysogenum leads to reduction of cephalosporin production and repression of nitrogen metabolism
Disruption of the nitrogen regulatory gene AcareA in Acremonium chrysogenum leads to reduction of cephalosporin production and repression of nitrogen metabolism
复制标题
产黄顶孢霉中氮调节基因 AcareA 的破坏导致头孢菌素产量减少并抑制氮代谢
DOI：
10.1016/j.fgb.2013.10.006
复制
发表时间：
2013-12-01
影响因子：
3
通讯作者：
Liu, Gang
中科院分区：
文献类型：
--
作者：
Li, Jinyang;Pan, Yuanyuan;Liu, Gang
AcareA基因是从产黄枝顶孢霉中克隆到的，编码真菌氮调节加塔锌指蛋白的同源物。基因破坏和遗传互补表明AcareA是氮代谢和头孢菌素生产所必需的。在以硝酸盐、尿酸和低浓度铵、谷氨酰胺或尿素为唯一氮源的培养基中，AcareA的破坏导致生长缺陷。转录分析表明，niaD/niiA的转录显着增加时，诱导硝酸盐在野生型和AcareA补充菌株，但不是在AcareA中断突变体。与头孢菌素生产的减少一致，编码头孢菌素生产酶的pcbAB、cefD 2、cefEF和cefG的转录在AcareA破坏突变体中减少。带移分析表明AcAREA与niaD、niA的启动子区和pcbAB-pcbC的双向启动子区结合。序列分析表明，所有的AcAREA结合位点都含有一致的加塔元件。这些结果表明，AcAREA在A.产金菌(C)2013 Elsevier Inc. All rights reserved.
AcareA, encoding a homologue of the fungal nitrogen regulatory GATA zinc-finger proteins, was cloned from Acremonium chrysogenum. Gene disruption and genetic complementation revealed that AcareA was required for nitrogen metabolism and cephalosporin production. Disruption of AcareA resulted in growth defect in the medium using nitrate, uric acid and low concentration of ammonium, glutamine or urea as sole nitrogen source. Transcriptional analysis showed that the transcription of niaD/niiA was increased drastically when induced with nitrate in the wild-type and AcareA complemented strains but not in AcareA disruption mutant. Consistent with the reduction of cephalosporin production, the transcription of pcbAB, cefD2, cefEF and cefG encoding the enzymes for cephalosporin production was reduced in AcareA disruption mutant. Band shift assays showed that AcAREA bound to the promoter regions of niaD, niiA and the bidirectional promoter region of pcbAB-pcbC. Sequence analysis showed that all the AcAREA binding sites contain the consensus GATA elements. These results indicated that AcAREA plays an important role both in the regulation of nitrogen metabolism and cephalosporin production in A. chrysogenum. (C) 2013 Elsevier Inc. All rights reserved.