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Characterization and expression of the Neurospora crassa nmt-1 gene

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Abstract

The Neurospora crassa homologue of the yeast no message in thiamine (nmt-1) gene was characterized. The deduced 342-amino-acid gene product has more than 60% identity with other fungal homologues and 42% similarity to a putative bacterial permease. In addition to three introns disrupting the coding sequence, a differentially spliced intron in the 5′ untranslated region was also detected. Unlike other fungi, the N. crassa nmt-1 gene is repressed only 6- to 8-fold by exogenous thiamine concentrations above 0.5 µM; and a high basal level of nmt-1 mRNA persists even at 5 µM thiamine. Immuno-blotting with purified antibodies detected two variants of NMT-1 which differ in size and charge. The more abundant 39-kDa form is more strongly repressed by thiamine than the 37-kDa protein. NMT-1 abundance modulates slowly in response to changes in the concentration of exogenous thiamine, suggesting that N. crassa maintains thiamine reserves in excess of immediate needs. Disruption of the nmt-1 gene demonstrated that it is essential for growth in the absence of exogenous thiamine. NMT-1-deficient strains had a growth rate and colony density which was about 70% of the wild type, despite supplementation with a wide range of exogenous thiamine. These results suggest that the nmt-1 gene plays some other role in addition to thiamine biosynthesis.

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Acknowledgements

This research was supported by a discovery grant from NSERC.

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Authors and Affiliations

  1. Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, Ontario, K1S 5B6, Canada

    Dorothy McColl, C. Alexander Valencia & P. John Vierula

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  1. Dorothy McColl
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  2. C. Alexander Valencia
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  3. P. John Vierula
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Correspondence to P. John Vierula.

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Communicated by U. Kück

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McColl, D., Valencia, C.A. & Vierula, P.J. Characterization and expression of the Neurospora crassa nmt-1 gene. Curr Genet 44, 216–223 (2003). https://doi.org/10.1007/s00294-003-0441-0

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