Characterization and expression of the Neurospora crassa nmt-1 gene
The Neurospora crassa homologue of the yeast no message in rtiiamine (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||Differential splicing, Growth, Thiamine|
McColl, D. (Dorothy), Valencia, C.A. (C. Alexander), & Vierula, J. (2003). Characterization and expression of the Neurospora crassa nmt-1 gene. Current Genetics, 44(4), 216–223. doi:10.1007/s00294-003-0441-0