Identification of the specific sequence recognized by Penicillium citrinum MlcR, a GAL4-type transcriptional activator of ML-236B (compactin) biosynthetic genes.
MlcR is a pathway-specific transcriptional activator of the ML-236B biosynthetic genes in Penicillium citrinum. The MlcR-binding sequences were identified by an in vitro gel-shift assay and an in vivo reporter assay for the region between mlcA and mlcC as a model. The gel-shift assay showed that recombinant MlcR bound to the DNA sequence 5'-ACGGCGTTATTCGG-3' and most of the bases in this motif were required for the interaction between MlcR and DNA. In the reporter assay using beta-glucuronidase (GUS), substitution of the bases in this binding sequence resulted in the drastic reduction of GUS activities. These data clearly indicate that this MlcR-binding sequence is essential for the transcriptional activation of mlcA and mlcC in P. citrinum. Similar motifs were found in other loci of the ML-236B biosynthetic gene cluster and the consensus-binding motif for MlcR was predicted to be a direct repeat, 5'-WCGG-N(6)-TCGG-3'.
Baba
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Process Technology Research Laboratories, Daiichi Sankyo Co., Ltd., 389-4 Aza-Ohtsurugi, Shimokawa, Izumi-machi, Iwaki, Fukushima 971-8183, Japan. satoshi_baba@icb.osaka-u.ac.jp
Nihira
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Hosobuchi
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D001483:Base Sequence; D001665:Binding Sites; D021901:DNA, Intergenic / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D008148:Lovastatin / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000235:genetics; D010407:Penicillium / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation
D021901:DNA, Intergenic; D005656:Fungal Proteins; D015534:Trans-Activators; C012258:mevastatin; D008148:Lovastatin