Impact of ammonium permeases mepA, mepB, and mepC on nitrogen-regulated secondary metabolism in Fusarium fujikuroi.
In Fusarium fujikuroi, the production of gibberellins and bikaverin is repressed by nitrogen sources such as glutamine or ammonium. Sensing and uptake of ammonium by specific permeases play key roles in nitrogen metabolism. Here, we describe the cloning of three ammonium permease genes, mepA, mepB, and mepC, and their participation in ammonium uptake and signal transduction in F. fujikuroi. The expression of all three genes is strictly regulated by the nitrogen regulator AreA. Severe growth defects of DeltamepB mutants on low-ammonium medium and methylamine uptake studies suggest that MepB functions as the main ammonium permease in F. fujikuroi. In DeltamepB mutants, nitrogen-regulated genes such as the gibberellin and bikaverin biosynthetic genes are derepressed in spite of high extracellular ammonium concentrations. mepA mepB and mepC mepB double mutants show a similar phenotype as DeltamepB mutants. All three F. fujikuroi mep genes fully complemented the Saccharomyces cerevisiae mep1 mep2 mep3 triple mutant to restore growth on low-ammonium medium, whereas only MepA and MepC restored pseudohyphal growth in the mep2/mep2 mutant. Overexpression of mepC in the DeltamepB mutants partially suppressed the growth defect but did not prevent derepression of AreA-regulated genes. These studies provide evidence that MepB functions as a regulatory element in a nitrogen sensing system in F. fujikuroi yet does not provide the sensor activity of Mep2 in yeast, indicating differences in the mechanisms by which nitrogen is sensed in S. cerevisiae and F. fujikuroi.
Teichert
Sabine
S
Westfälische Wilhelms-Universität Münster, Institut für Botanik, Schlossgarten 3, D-48149 Münster, Germany.
Rutherford
Julian C
JC
Wottawa
Marieke
M
Heitman
Joseph
J
Tudzynski
Bettina
B
D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology*; D005670:Fusarium / Q000254:growth & development / Q000378:metabolism* / Q000472:pathogenicity; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D005816:Genetic Complementation Test; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D009584:Nitrogen / Q000494:pharmacology*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D000644:Quaternary Ammonium Compounds / Q000378:metabolism*; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism; D014170:Transformation, Genetic
D005656:Fungal Proteins; D026901:Membrane Transport Proteins; D000644:Quaternary Ammonium Compounds; D012331:RNA, Fungal; D012333:RNA, Messenger; D009584:Nitrogen