Figure 1: a, Unrooted phylogenetic tree derived from full-length amino acid sequence alignments showing the relationship of G. sulfurreducens pilin domain.

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Figure 1: a, Unrooted phylogenetic tree derived from full-length amino acid sequence alignments showing the relationship of G. sulfurreducens pilin domain proteins, GSU1496 and GSU1776, and pilin and pseudopilin proteins from other bacteria. The tree was obtained by the neighbor-joining method using the BioNJ algorithm . Bootstrap values were determined from 100 replicates. GSU1496, Geobacter sulfurreducens (gi:399996596); PilA, Geobacter metallireducens (gi:23053459); GSU1776, G. sulfurreducens (gi:41152675); GspG, G. sulfurreducens (gi:399995435); PilA, Deinococcus radiodurans (gi:15805575); PilA, Vibrio cholerae El Tor N16961 (gi:4455001); PilA, Pseudomonas aeruginosa PA302025 (gi:21629635); PilA, Pseudomonas fluorescens PfO-1 (gi:23058745); NE1308, Nitrosomonas europaea ATCC 19718 (gi:30249287); PilE, Neisseria meningiditis FAM18 (gi:2228578); GspG, Mesorhizobium loti MAF 303099 (gi:13475692); PilA, P. aeruginosa PA5196 (gi:21464534); EspG, V. cholerae El Tor N16961 (gi:15642724); PilA, Ralstonia metallidurans (gi:15642724); GspG, Burkholderia cepapcia KF1 (gi:22978954); PilA, Dichelobacter nodosus N2 (gi:16118465); PulG, P. fluorescens PfO-1 (gi:23059410); PilE, Neisseria gonorrhoeae FA1090 (gi:2921589); GspG, Thermoanaeobacter tengcongensis MB4T (gi:20807714); HofG, Synechocystis sp. PCC6803 (gi:16330343); Yts1G, Yersinia enterocolitica WA-314 (gi:27529239). b, Genomic organization of pilus biosynthesis genes surrounding ORF GSU1496, a pilA homologue of G. sulfurreducens (top). This arrangement is analogous to the pil genes of Myxococcus xanthus (bottom). A PilA- mutant strain was constructed by replacement of the central portion of the gene with a chloramphenicol cassette. Figure S1 Amino acid sequence alignment of pilin domain proteins (GSU1496 (PilA) and GSU1776 (OxpG)) from G. sulfurreducens and pilins and pseudopilins from other bacteria. Amino acid sequence alignments were first generated using the Clustal X (1.8) program and manually optimized using the SeAl v. 2.0a11 Carbon program (University of Oxford). The alignment contains the conserved amino-terminal domain of pilins and pseudopilins and was used to infer the phylogenetic tree shown in Fig. 1c. Different amino acid residues were assigned to one of a number of groups (by chemical similarity) and each group was assigned a colour.

5 10 15 20 25 30 Fe(II) (mM) wild-type PilA- mutant FeOOH + NTA +AQDS Figure 1: a, Unrooted phylogenetic tree derived from full-length amino acid sequence alignments showing the relationship of G. sulfurreducens pilin domain proteins, GSU1496 and GSU1776, and pilin and pseudopilin proteins from other bacteria. The tree was obtained by the neighbor-joining method using the BioNJ algorithm . Bootstrap values were determined from 100 replicates. GSU1496, Geobacter sulfurreducens (gi:399996596); PilA, Geobacter metallireducens (gi:23053459); GSU1776, G. sulfurreducens (gi:41152675); GspG, G. sulfurreducens (gi:399995435); PilA, Deinococcus radiodurans (gi:15805575); PilA, Vibrio cholerae El Tor N16961 (gi:4455001); PilA, Pseudomonas aeruginosa PA302025 (gi:21629635); PilA, Pseudomonas fluorescens PfO-1 (gi:23058745); NE1308, Nitrosomonas europaea ATCC 19718 (gi:30249287); PilE, Neisseria meningiditis FAM18 (gi:2228578); GspG, Mesorhizobium loti MAF 303099 (gi:13475692); PilA, P. aeruginosa PA5196 (gi:21464534); EspG, V. cholerae El Tor N16961 (gi:15642724); PilA, Ralstonia metallidurans (gi:15642724); GspG, Burkholderia cepapcia KF1 (gi:22978954); PilA, Dichelobacter nodosus N2 (gi:16118465); PulG, P. fluorescens PfO-1 (gi:23059410); PilE, Neisseria gonorrhoeae FA1090 (gi:2921589); GspG, Thermoanaeobacter tengcongensis MB4T (gi:20807714); HofG, Synechocystis sp. PCC6803 (gi:16330343); Yts1G, Yersinia enterocolitica WA-314 (gi:27529239). b, Genomic organization of pilus biosynthesis genes surrounding ORF GSU1496, a pilA homologue of G. sulfurreducens (top). This arrangement is analogous to the pil genes of Myxococcus xanthus (bottom). A PilA- mutant strain was constructed by replacement of the central portion of the gene with a chloramphenicol cassette. Figure S2 Restoration of Fe(III) oxide (FeOOH) reduction by the PilA- mutant after media supplementation with 4 mM NTA, an Fe(III) chelator, or 0.1 mM AQDS, an electron shuttle that alleviates the need for establishing contact with the insoluble Fe(III) oxides. The levels of soluble Fe(II), produced as a result of the reduction and growth on Fe(III) oxides, were measured after 3 weeks of incubation with the insoluble electron acceptor.

a b 0.1 µm 0.1 µm 10 nm c Figure 1: a, Unrooted phylogenetic tree derived from full-length amino acid sequence alignments showing the relationship of G. sulfurreducens pilin domain proteins, GSU1496 and GSU1776, and pilin and pseudopilin proteins from other bacteria. The tree was obtained by the neighbor-joining method using the BioNJ algorithm . Bootstrap values were determined from 100 replicates. GSU1496, Geobacter sulfurreducens (gi:399996596); PilA, Geobacter metallireducens (gi:23053459); GSU1776, G. sulfurreducens (gi:41152675); GspG, G. sulfurreducens (gi:399995435); PilA, Deinococcus radiodurans (gi:15805575); PilA, Vibrio cholerae El Tor N16961 (gi:4455001); PilA, Pseudomonas aeruginosa PA302025 (gi:21629635); PilA, Pseudomonas fluorescens PfO-1 (gi:23058745); NE1308, Nitrosomonas europaea ATCC 19718 (gi:30249287); PilE, Neisseria meningiditis FAM18 (gi:2228578); GspG, Mesorhizobium loti MAF 303099 (gi:13475692); PilA, P. aeruginosa PA5196 (gi:21464534); EspG, V. cholerae El Tor N16961 (gi:15642724); PilA, Ralstonia metallidurans (gi:15642724); GspG, Burkholderia cepapcia KF1 (gi:22978954); PilA, Dichelobacter nodosus N2 (gi:16118465); PulG, P. fluorescens PfO-1 (gi:23059410); PilE, Neisseria gonorrhoeae FA1090 (gi:2921589); GspG, Thermoanaeobacter tengcongensis MB4T (gi:20807714); HofG, Synechocystis sp. PCC6803 (gi:16330343); Yts1G, Yersinia enterocolitica WA-314 (gi:27529239). b, Genomic organization of pilus biosynthesis genes surrounding ORF GSU1496, a pilA homologue of G. sulfurreducens (top). This arrangement is analogous to the pil genes of Myxococcus xanthus (bottom). A PilA- mutant strain was constructed by replacement of the central portion of the gene with a chloramphenicol cassette. Figure S3 TEM to negatively stained wild-type pili. (a-b) G. sulfurreducens cells expressing pili after 48h of incubation at 250C in medium with fumarate. (c) Pili filaments that have been mechanically sheared off the outer surface of the wild-type strain of G. sulfurreducens. The wild-type pili preparations were analyzed by conducting-probe AFM (Fig. 4) in order to study the electroconductive properties of G. sulfurreducens pili.

100 nm 20 nm a 1 nA b c 100 mV Figure 1: a, Unrooted phylogenetic tree derived from full-length amino acid sequence alignments showing the relationship of G. sulfurreducens pilin domain proteins, GSU1496 and GSU1776, and pilin and pseudopilin proteins from other bacteria. The tree was obtained by the neighbor-joining method using the BioNJ algorithm . Bootstrap values were determined from 100 replicates. GSU1496, Geobacter sulfurreducens (gi:399996596); PilA, Geobacter metallireducens (gi:23053459); GSU1776, G. sulfurreducens (gi:41152675); GspG, G. sulfurreducens (gi:399995435); PilA, Deinococcus radiodurans (gi:15805575); PilA, Vibrio cholerae El Tor N16961 (gi:4455001); PilA, Pseudomonas aeruginosa PA302025 (gi:21629635); PilA, Pseudomonas fluorescens PfO-1 (gi:23058745); NE1308, Nitrosomonas europaea ATCC 19718 (gi:30249287); PilE, Neisseria meningiditis FAM18 (gi:2228578); GspG, Mesorhizobium loti MAF 303099 (gi:13475692); PilA, P. aeruginosa PA5196 (gi:21464534); EspG, V. cholerae El Tor N16961 (gi:15642724); PilA, Ralstonia metallidurans (gi:15642724); GspG, Burkholderia cepapcia KF1 (gi:22978954); PilA, Dichelobacter nodosus N2 (gi:16118465); PulG, P. fluorescens PfO-1 (gi:23059410); PilE, Neisseria gonorrhoeae FA1090 (gi:2921589); GspG, Thermoanaeobacter tengcongensis MB4T (gi:20807714); HofG, Synechocystis sp. PCC6803 (gi:16330343); Yts1G, Yersinia enterocolitica WA-314 (gi:27529239). b, Genomic organization of pilus biosynthesis genes surrounding ORF GSU1496, a pilA homologue of G. sulfurreducens (top). This arrangement is analogous to the pil genes of Myxococcus xanthus (bottom). A PilA- mutant strain was constructed by replacement of the central portion of the gene with a chloramphenicol cassette. Figure S4 CP-AFM analyses to Shewanella oneidensis MR-1 pili. (a) Height image of a pilus filament. (b) Current map obtained while applying a sweeping voltage (c) showing no detectable conductivity across the pilus filament.

100 nm 15 nm a 1 nA b c 200 mV Figure 1: a, Unrooted phylogenetic tree derived from full-length amino acid sequence alignments showing the relationship of G. sulfurreducens pilin domain proteins, GSU1496 and GSU1776, and pilin and pseudopilin proteins from other bacteria. The tree was obtained by the neighbor-joining method using the BioNJ algorithm . Bootstrap values were determined from 100 replicates. GSU1496, Geobacter sulfurreducens (gi:399996596); PilA, Geobacter metallireducens (gi:23053459); GSU1776, G. sulfurreducens (gi:41152675); GspG, G. sulfurreducens (gi:399995435); PilA, Deinococcus radiodurans (gi:15805575); PilA, Vibrio cholerae El Tor N16961 (gi:4455001); PilA, Pseudomonas aeruginosa PA302025 (gi:21629635); PilA, Pseudomonas fluorescens PfO-1 (gi:23058745); NE1308, Nitrosomonas europaea ATCC 19718 (gi:30249287); PilE, Neisseria meningiditis FAM18 (gi:2228578); GspG, Mesorhizobium loti MAF 303099 (gi:13475692); PilA, P. aeruginosa PA5196 (gi:21464534); EspG, V. cholerae El Tor N16961 (gi:15642724); PilA, Ralstonia metallidurans (gi:15642724); GspG, Burkholderia cepapcia KF1 (gi:22978954); PilA, Dichelobacter nodosus N2 (gi:16118465); PulG, P. fluorescens PfO-1 (gi:23059410); PilE, Neisseria gonorrhoeae FA1090 (gi:2921589); GspG, Thermoanaeobacter tengcongensis MB4T (gi:20807714); HofG, Synechocystis sp. PCC6803 (gi:16330343); Yts1G, Yersinia enterocolitica WA-314 (gi:27529239). b, Genomic organization of pilus biosynthesis genes surrounding ORF GSU1496, a pilA homologue of G. sulfurreducens (top). This arrangement is analogous to the pil genes of Myxococcus xanthus (bottom). A PilA- mutant strain was constructed by replacement of the central portion of the gene with a chloramphenicol cassette. Figure S5 CP-AFM analyses to Pseudomonas aeruginosa strain K pili. (a) Height image of several pilus filaments. (b) Current map obtained while applying a sweeping voltage (c) showing no detectable conductivity across the pilus filament.