Tat-Dependent Translocation of an F420-Binding Protein of Mycobacterium tuberculosis

F-420 is a unique cofactor present in a restricted range of microorganisms, including mycobacteria. It has been proposed that F-420 has an important role in the oxidoreductive reactions of Mycobacterium tuberculosis, possibly associated with anaerobic survival and persistence. The protein encoded by Rv0132c has a predicted N-terminal signal sequence and is annotated as an F-420-dependent glucose-6-phosphate dehydrogenase. Here we show that Rv0132c protein does not have the annotated activity. It does, however, co-purify with F-420 during expression experiments in M. smegmatis. We also show that the Rv0132c-F-420 complex is a substrate for the Tat pathway, which mediates translocation of the complex across the cytoplasmic membrane, where Rv0132c is anchored to the cell envelope. This is the first report of any F-420-binding protein being a substrate for the Tat pathway and of the presence of F-420 outside of the cytosol in any F-420-producing microorganism. The Rv0132c protein and its Tat export sequence are essentially invariant in the Mycobacterium tuberculosis complex. Taken together, these results show that current understanding of F-420 biology in mycobacteria should be expanded to include activities occurring in the extra-cytoplasmic cell envelope.