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Current Genetics

, Volume 44, Issue 4, pp 216–223 | Cite as

Characterization and expression of the Neurospora crassa nmt-1 gene

  • Dorothy McColl
  • C. Alexander Valencia
  • P. John Vierula
Research Article

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.

Keywords

Thiamine Differential splicing Growth 

Notes

Acknowledgements

This research was supported by a discovery grant from NSERC.

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Dorothy McColl
    • 1
  • C. Alexander Valencia
    • 1
  • P. John Vierula
    • 1
  1. 1.Department of BiologyCarleton UniversityOttawaCanada

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