CebR as a master regulator for cellulose/cellooligosaccharide catabolism affects morphological development in Streptomyces griseus.
Streptomyces griseus mutants exhibiting deficient glucose repression of beta-galactosidase activity on lactose-containing minimal medium supplemented with a high concentration of glucose were isolated. One of these mutants had a 12-bp deletion in cebR, which encodes a LacI/GalR family regulator. Disruption of cebR in the wild-type strain caused the same phenotype as the mutant, indicating that CebR is required for glucose repression of beta-galactosidase activity. Recombinant CebR protein bound to a 14-bp inverted-repeat sequence (designated the CebR box) present in the promoter regions of cebR and the putative cellobiose utilization operon, cebEFG-bglC. The DNA-binding activity of CebR was impaired by cellooligosaccharides, including cellobiose, cellotriose, cellotetraose, cellopentaose, and cellohexaose. In agreement with this observation, transcription from the cebE and cebR promoters was greatly enhanced by the addition of cellobiose to the medium. Seven other genes containing one or two CebR boxes in their upstream regions were found in the S. griseus genome. Five of these genes encode putative secreted proteins: two cellulases, a cellulose-binding protein, a pectate lyase, and a protein of unknown function. These five genes and cebEFG-bglC were transcribed at levels 4 to 130 times higher in the DeltacebR mutant than in the wild-type strain, as determined by quantitative reverse transcription-PCR. These findings indicate that CebR is a master regulator of cellulose/cellooligosaccharide catabolism. Unexpectedly, the DeltacebR mutant formed very few aerial hyphae on lactose-containing medium, demonstrating a link between carbon source utilization and morphological development.
Marushima
Kazuya
K
Department of Biotechnology, Graduate School of Agriculture and Life Sciences, The University of Tokyo, Tokyo, Japan.
Ohnishi
Yasuo
Y
Horinouchi
Sueharu
S
D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D002475:Cellobiose / Q000378:metabolism; D002482:Cellulose / Q000031:analogs & derivatives / Q000378:metabolism*; D004247:DNA / Q000235:genetics / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics / Q000502:physiology; D005816:Genetic Complementation Test; D009154:Mutation; D009844:Oligosaccharides / Q000378:metabolism*; D009876:Operon / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013305:Streptomyces griseus / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D013780:Tetroses / Q000378:metabolism
D001426:Bacterial Proteins; D009844:Oligosaccharides; D013780:Tetroses; D002475:Cellobiose; C099423:cellohexaose; C019193:maltopentaose; C048468:cellotetraose; D002482:Cellulose; D004247:DNA