Fe-hydrogenase maturases in the hydrogenosomes of Trichomonas vaginalis.
Assembly of active Fe-hydrogenase in the chloroplasts of the green alga Chlamydomonas reinhardtii requires auxiliary maturases, the S-adenosylmethionine-dependent enzymes HydG and HydE and the GTPase HydF. Genes encoding homologous maturases had been found in the genomes of all eubacteria that contain Fe-hydrogenase genes but not yet in any other eukaryote. By means of proteomic analysis, we identified a homologue of HydG in the hydrogenosomes, mitochondrion-related organelles that produce hydrogen under anaerobiosis by the activity of Fe-hydrogenase, in the pathogenic protist Trichomonas vaginalis. Genes encoding two other components of the Hyd system, HydE and HydF, were found in the T. vaginalis genome database. Overexpression of HydG, HydE, and HydF in trichomonads showed that all three proteins are specifically targeted to the hydrogenosomes, the site of Fe-hydrogenase maturation. The results of Neighbor-Net analyses of sequence similarities are consistent with a common eubacterial ancestor of HydG, HydE, and HydF in T. vaginalis and C. reinhardtii, supporting a monophyletic origin of Fe-hydrogenase maturases in the two eukaryotes. Although Fe-hydrogenases exist in only a few eukaryotes, related Narf proteins with different cellular functions are widely distributed. Thus, we propose that the acquisition of Fe-hydrogenases, together with Hyd maturases, occurred once in eukaryotic evolution, followed by the appearance of Narf through gene duplication of the Fe-hydrogenase gene and subsequent loss of the Hyd proteins in eukaryotes in which Fe-hydrogenase function was lost.
Pütz
Simone
S
Institut für Botanik III, Universitätsstrasse 1, 40225 Düsseldorf, Germany.
Dolezal
Pavel
P
Gelius-Dietrich
Gabriel
G
Bohacova
Lenka
L
Tachezy
Jan
J
Henze
Katrin
K
D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002458:Cell Fractionation; D017124:Conserved Sequence; D015180:Electrophoresis, Gel, Two-Dimensional; D005455:Fluorescent Antibody Technique; D017125:Genes, Protozoan; D016678:Genome; D006864:Hydrogenase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007506:Iron-Sulfur Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007526:Isoelectric Point; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D016366:Open Reading Frames; D010802:Phylogeny; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D014170:Transformation, Genetic; D014246:Trichomonas vaginalis / Q000166:cytology / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development
D007506:Iron-Sulfur Proteins; D015800:Protozoan Proteins; C118830:iron hydrogenase; D006864:Hydrogenase