Figure S1. Phylogenetic tree comparing c-type cytochrome proteins from Ferroglobus placidus to c-type cytochromes from other bacterial and archaeal

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1 Fiure S1. Phyloenetic tree comparin c-type cytochrome proteins from Ferrolobus placidus to c-type cytochromes from other bacterial and archaeal species. 1

2 Fiure S2. Alternative complex III operons found in Ferrolobus placidus (locus id: Ferp), Geobacter bemidjiensis (Gbem), G. metallireducens (Gmet), G. daltonii (Geob), G. sulfurreducens (GSU), G. uraniireducnes (Gura), G. bremensis (K419DRAFT), strain M18 (GM18), strain M21 (GM21), Geopsychrobacter electrodiphilus (D888DRAFT), Anaeromyxobacter sp. FW109-5 (Anae109), A. dehaloenans (Adeh), Desulfocapsa sulfexiens (UWK), and Desulfobacula toluolica (TOL2). 2

3 Table S1. Primers used to quantify different mrna transcripts from F. placidus by quantitative RT-PCR. Locus ID Ferp_0660 Ferp_0668 Ferp_0670 Ferp_0672 Ferp_1255 Ferp_1267 Ferp_1336 Ferp_1341 Ferp_1813 Ferp_1814 Ferp_2064 Primer name Ferp_ f/336r Ferp_ f/550r Ferp_ f/2184r Ferp_ f/4410r Ferp_ f/313r Ferp_ f/434r Ferp_ f/613r Ferp_ f/619r Ferp_ f/309r Ferp_ f/256r Ferp_ f/537r Forward primer (5 3 ) Reverse primer (5 3 ) TTACACGGAGTCGACGTTTG CAGCTTGTAGTGGGGATCGT CACAGACCGTGGGTTCTTTCA CGTGGGTGTGACAGCCTATG CATCAGGTTCGGAAGGGACATA GGCATAGTGGGAGACCGTAGAG CGCCGACAATCTTTCAATACTG CCCGTTTACTGCCCTGTTCTC GACGTTGGCAACAACTACGA AGAGGACGTCTTTGGCGTAA ATTCTGCCTGAACTGCCACAA CAAACCGTACAAGGCTTATCCG CAAAATACAAGCCGCCGAAA CGGATGCTTTACACCAGTTGG CGATTACAGTTTTGCGACGA CGAGGAAGACCGTGAAGAAG CGCCAACACCAAAACCTACT GTTACCGCAGAGCTGTCCTC CAACAATCCGAAGAGGCAAT GGCAGGAAATGCAGTAATCG ATGCCATAAACGGATTGAGACA GGCGAAGCACCAGAAGTTG 3

4 Table S2. Evaluation of c-type cytochromes present in the enomes of hyperthermophilic archaea. Hyperthermophilic archaeon Archaeolobus fulidus Geolobus acetivorans Caldivira maquilinensis Ferrolobus placidus Methanopyrus kandleri Pyrobaculum aerophilum Pyrobaculum islandicum Tempera ture rane or optimum ( C) Total putat ive cyt c enes with 1 motif with 2-5 motifs with 6-19 motifs with 20+ motifs Electron acceptors utilized Fe(III), SO 2-4, SO 2-3, S 2 O 2-3 (1-3) Fe(III) (4, 5) SO 2-4, SO 2-3, S 2 O 2-3, NO - 3, Fe(III) (6) Fe(III), S 2 O 2-3, NO - 3 (7, 8) Fe(III), CO 2 (3, 9) O 2, NO - 3, NO - 2, Fe(III) (10, 11) SO 2-4, SO 2-3, S 2 O 2-3, Fe(III), Mn(IV), U(VI), Tc(VII), Cr(VI), Co(III) (12, 13) NO - 3, AsO 3-4, S 2 O 2-3, Fe(III) (14) Pyrobaculum sp Pyrococcus abyssi fermentative, Fe(III) (3, 15) Pyrococcus furiosus fermentative, Fe(III) (3, 16, 17) 4

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6 6. Itoh T, Suzuki K, Sanchez PC, Nakase T Caldivira maquilinensis en. nov., sp nov., a new enus of rod-shaped crenarchaeote isolated from a hot sprin in the Philippines. Int. J. Syst. Bacteriol. 49: Hafenbradl D, Keller M, Dirmeier R, Rachel R, Rossnael P, Burraf S, Huber H, Stetter KO Ferrolobus placidus en nov, sp nov, a novel hyperthermophilic archaeum that oxidizes Fe 2+ at neutral ph under anoxic conditions. Arch. Microbiol. 166: Tor JM, Kashefi K, Lovley DR Acetate oxidation coupled to Fe(III) reduction in hyperthermophilic microoranisms. Appl. Environ. Microb. 67: Kurr M, Huber R, Koni H, Jannasch HW, Fricke H, Trincone A, Kristjansson JK, Stetter KO Methanopyrus kandleri, Gen and Sp-nov represents a novel roup of hyperthermophilic methanoens, rowin at 110-derees-C. Arch. Microbiol. 156: Volkl P, Huber R, Drobner E, Rachel R, Burraf S, Trincone A, Stetter KO Pyrobaculum aerophilum sp-nov, a novel nitrate-reducin hyperthermophilic archaeum. Appl. Environ. Microb. 59: Feinber LF, Holden JF Characterization of dissimilatory Fe(III) versus NO 3 - reduction in the hyperthermophilic archaeon Pyrobaculum aerophilum. J. Bacteriol. 188: Huber R, Kristjansson JK, Stetter KO Pyrobaculum en-nov, a new enus of neutrophilic, rod-shaped Archaebacteria from continental solfataras rowin optimally at 100-Derees-C. Arch. Microbiol. 149:

7 13. Kashefi K, Lovley DR Reduction of Fe(III), Mn(IV), and toxic metals at 100 derees C by Pyrobaculum islandicum. Appl. Environ. Microb. 66: Mardanov AV, Gumerov VM, Slobodkina GB, Beletsky AV, Bonch-Osmolovskaya EA, Ravin NV, Skryabina KG Complete enome sequence of strain 1860, a crenarchaeon of the enus Pyrobaculum able to row with various electron acceptors. J. Bacteriol. 194: Erauso G, Reysenbach AL, Godfroy A, Meunier JR, Crump B, Partensky F, Baross JA, Marteinsson V, Barbier G, Pace NR, Prieur D Pyrococcus abyssi sp-nov, a new hyperthermophilic archaeon isolated from a deep-sea hydrothermal vent. Arch. Microbiol. 160: Fiala G, Stetter KO Pyrococcus furiosus sp-nov represents a novel enus of marine heterotrophic Archaebacteria rowin optimally at 100-Derees C. Arch. Microbiol. 145: Robb FT, Maeder DL, Brown JR, DiRuiero J, Stump MD, Yeh RK, Weiss RB, Dunn DM Genomic sequence of hyperthermophile, Pyrococcus furiosus: implications for physioloy and enzymoloy. Methods Enzymol. 330: