Tagging of a nitrogen pathway-specific regulator gene in Tolypocladium inflatum by the transposon Restless.
Restless is an endogenous hAT transposon found in the cyclosporin-producing fungus Tolypocladium inflatum. This element is present in about 15 copies in a particular strain (ATCC34921) which was used for successful gene tagging. We have isolated a T. inflatum mutant with a defect in nitrogen metabolism. This mutant carries a copy of the Restless element in a gene encoding a C6 zinc-finger protein. The deduced amino acid sequence of the gene product shows a significant similarity to the NIT4 protein of Neurospora crassa, which is a regulator of nitrogen metabolism. The wild-type T. inflatum gene was shown to complement a nit-4 mutant of N. crassa. From these data, we conclude that the T. inflatum gene also encodes a regulator of nitrogen metabolism, which was named tnir1 (Tolypocladium nitrogen regulator 1). To the best of our knowledge, this is the first fungal gene to be identified by transposon-directed gene tagging. A general method for gene tagging using an endogenous fungal transposon is presented.
Kempken
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F
Lehrstuhl für Allgemeine und Molekulare Botanik, Ruhr-Universität Bochum, Germany.
Kück
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D000595:Amino Acid Sequence; D001203:Ascomycota / Q000235:genetics*; D003001:Cloning, Molecular; D004251:DNA Transposable Elements / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics; D005656:Fungal Proteins / Q000378:metabolism*; D015723:Gene Library; D005809:Genes, Regulator; D005816:Genetic Complementation Test; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009492:Neurospora crassa / Q000235:genetics; D009584:Nitrogen / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators; D014158:Transcription, Genetic
D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D015534:Trans-Activators; C071240:NIT-4 protein, Neurospora crassa; D009584:Nitrogen