title: Disruption of the nitrogen regulatory gene AcareA in Acremonium chrysogenum leads to reduction of cephalosporin production and repression of nitrogen metabolism.
abstract: 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.
authors.affiliation: State Key Laboratory of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Jinyang
authors.initials: J
authors.lastname: Li
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Pan
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
mesh_terms: D000164:Acremonium / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D002511:Cephalosporins / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D020869:Gene Expression Profiling; D055786:Gene Knockout Techniques; D005809:Genes, Regulator; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017422:Sequence Analysis, DNA; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D000900:Anti-Bacterial Agents; D002511:Cephalosporins; D003470:Culture Media; D004271:DNA, Fungal; D014157:Transcription Factors; D009584:Nitrogen
keywords: AcareA; Acremonium chrysogenum; Cephalosporin; Nitrogen metabolism