Fig. 2

Mitochondrial Tat proteins of A. godoyi but substitute for the function of E. coli TAT pathway. a Diagrams depict the principle of the complementation assay aimed at the translocation of TAT substrate (AmiA) across the inner E. coli membrane. + (pTAT101 plasmid carrying complete tat operon) and − (pTAT101 without tat operon) represent the positive and negative controls, respectively. In all strains (MCDSSAC Δtat), mature domain of AmiA with SufI Tat signal peptide was expressed. While A. godoyi mitochondrial TatA and TatC (expressed from pTAT101) could functionally replace E. coli proteins, N. gruberi TatC could not rescue the function when expressed in a synthetic operon with E. coli proteins. b Individual TatA and TatC were expressed from plasmids containing promotor of varying strength (tat or T7 promotor) in corresponding E. coli mutant (top line of each panel). None of the gene could functionally replace the bacterial protein or, in case of TatC, the whole TAT system. Ag—A. godoyi, Ng—N. gruberi, Ra—R. americana, Mj—M. jakobiformis. c Diagrams depict the clustering of YFP-tagged TatA (TatAy) induced upon the overexpression TAT substrate (CueO). Replacing E. coli TatC with NgTatC abolished the formation of the active translocase clusters