Genetic and molecular characterization reveals a unique nucleobase cation symporter 1 in Arabidopsis

► Here we identified the sole nucleobase cation symporter 1 (AtNCS1) in Arabidopsis. ► In planta and heterologous expression studies revealed solute specificity of AtNCS1. ► AtNCS1 is unique in moving adenine (A), guanine (G), and uracil (U). ► This is in contrast to yeast’s NCS1 FUR4 (transports U) or FCY2 (transports A, G, and C). Locus At5g03555 encodes a nucleobase cation symporter 1 (AtNCS1) in the Arabidopsis genome. Arabidopsis insertion mutants, AtNcs1-1 and AtNcs1-3, were used for in planta toxic nucleobase analog growth studies and radio-labeled nucleobase uptake assays to characterize solute transport specificities. These results correlate with similar growth and uptake studies of AtNCS1 expressed in Saccharomyces cerevisiae. Both in planta and heterologous expression studies in yeast revealed a unique solute transport profile for AtNCS1 in moving adenine, guanine and uracil. This is in stark contrast to the canonical transport profiles determined for the well-characterized S. cerevisiae NCS1 proteins FUR4 (uracil transport) or FCY2 (adenine, guanine, and cytosine transport).