Accessory proteins functioning selectively and pleiotropically in the biosynthesis of [NiFe] hydrogenases in Thiocapsa roseopersicina.
There are at least two membrane-bound (HynSL and HupSL) and one soluble (HoxEFUYH) [NiFe] hydrogenases in Thiocapsa roseopersicina BBS, a purple sulfur photosynthetic bacterium. Genes coding for accessory proteins that participate in the biosynthesis and maturation of hydrogenases seem to be scattered along the chromosome. Transposon-based mutagenesis was used to locate the hydrogenase accessory genes. Molecular analysis of strains showing mutant phenotypes led to the identification of hupK (hoxV ), hypC1, hypC2, hypD, hypE, and hynD genes. The roles of hynD, hupK and the two hypC genes were investigated in detail. The putative HynD was found to be a hydrogenase-specific endoprotease type protein, participating in the maturation of the HynSL enzyme. HupK plays an important role in the formation of the functionally active membrane-bound [NiFe] hydrogenases, but not in the biosynthesis of the soluble enzyme. In-frame deletion mutagenesis showed that HypC proteins were not specific for the maturation of either hydrogenase enzyme. The lack of either HypC protein drastically reduced the activity of every hydrogenase. Hence both HypCs might participate in the maturation of [NiFe] hydrogenases. Homologous complementation with the appropriate genes substantiated the physiological roles of the corresponding gene products in the H2 metabolism of T. roseopersicina.
Maróti
Gergely
G
Institute of Biophysics, Biological Research Center, Hungarian Academy of Sciences, Hungary.
Fodor
Barna D
BD
Rákhely
Gábor
G
Kovács
Akos T
AT
Arvani
Solmaz
S
Kovács
Kornél L
KL
D001426:Bacterial Proteins / Q000235:genetics; D015139:Blotting, Southern; D002462:Cell Membrane / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D004251:DNA Transposable Elements; D010450:Endopeptidases / Q000737:chemistry; D017353:Gene Deletion; D005816:Genetic Complementation Test; D006859:Hydrogen / Q000737:chemistry; D006864:Hydrogenase / Q000737:chemistry* / Q000378:metabolism*; D008957:Models, Genetic; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011506:Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D020586:Thiocapsa / Q000201:enzymology*; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; C108874:HypC protein, Bacteria; C108875:HypD protein, Bacteria; D011506:Proteins; C089912:hupK protein, Rhizobium leguminosarum; C085871:hypE protein, Bacteria; D006859:Hydrogen; D004247:DNA; C043296:nickel-iron hydrogenase; D006864:Hydrogenase; D010450:Endopeptidases