A UPF0118 family protein with uncharacterized function from the moderate halophile Halobacillus andaensis represents a novel class of Na(+)(Li(+))/H(+) antiporter

In this study, genomic DNA was screened from Halobacillus andaensis NEAU-ST10-40(T) by selection in Escherichia coli KNabc lacking three major Na(+)/H(+) antiporters. One gene designated upf0118 exhibiting Na(+)(Li(+))/H(+) antiport activity was finally cloned. Protein alignment showed that UPF0118 shares the highest identity of 81.5% with an unannotated gene encoding a protein with uncharacterized protein function belonging to UPF0118 family from H. kuroshimensis, but shares no identity with all known specific Na(+)(Li(+))/H(+) antiporter genes or genes with Na(+)(Li(+))/H(+) antiport activity. Growth test, western blot and Na(+)(Li(+))/H(+) antiport assay revealed that UPF0118 as a transmembrane protein exhibits pH-dependent Na(+)(Li(+))/H(+) antiport activity. Phylogenetic analysis indicated that UPF0118 clustered with all its homologs belonging to UPF0118 family at a wide range of 22–82% identities with the bootstrap value of 92%, which was significantly distant with all known specific single-gene Na(+)(Li(+))/H(+) antiporters and single-gene proteins with the Na(+)(Li(+))/H(+) antiport activity. Taken together, we propose that UPF0118 should represent a novel class of Na(+)(Li(+))/H(+) antiporter. To the best of our knowledge, this is the first report on the functional analysis of a protein with uncharacterized protein function as a representative of UPF0118 family containing the domain of unknown function, DUF20.