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

摘要

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.