Missense mutations that inactivate the Aspergillus nidulans nrtA gene encoding a high-affinity nitrate transporter

Kinghorn, J, Sloan, J, Kana'a, G, DeSilva, E, Rouch, D and Unkles, S 2005, 'Missense mutations that inactivate the Aspergillus nidulans nrtA gene encoding a high-affinity nitrate transporter', Genetics, vol. 169, no. 3, pp. 1369-1377.


Document type: Journal Article
Collection: Journal Articles

Title Missense mutations that inactivate the Aspergillus nidulans nrtA gene encoding a high-affinity nitrate transporter
Author(s) Kinghorn, J
Sloan, J
Kana'a, G
DeSilva, E
Rouch, D
Unkles, S
Year 2005
Journal name Genetics
Volume number 169
Issue number 3
Start page 1369
End page 1377
Total pages 9
Publisher Genetics Society of America
Abstract The transport of nitrate into prokaryotic and eukaryotic cells, of considerable interest to agriculture, ecology, and human health, is carried out by members of a distinct cluster of proteins within the major facilitator superfamily. To obtain structure/function information on this important class of nitrate permeases, a collection of chemically induced mutations in the nrtA gene encoding a 12-transmembrane domain, high-affinity nitrate transporter from the eukaryote Aspergillus nidulans was isolated and characterized. This mutational analysis, coupled with protein alignments, demonstrates the utility of the approach to predicting peptide motifs and individual residues important for the movement of nitrate across the membrane. These include the highly conserved nitrate signature motif (residues 166-173) in Tm 5, the conserved charged residues Arg87 (Tm 2) and Arg368 (Tm 8), as well as the aromatic residue Phe47 (Tm 1), all within transmembrane helices. No mutations were observed in the large central loop (Lp 6/7) between Tm 6 and Tm 7. Finally, the study of a strain with a conversion of Trp481 (Tm 12) to a stop codon suggests that all 12 transmembrane domains and/or the C-terminal tail are required for membrane insertion and/or stability of NrtA.
Subject Receptors and Membrane Biology
DOI - identifier 10.1534/genetics.104.036590
ISSN 0016-6731
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