Rhodobacter capsulatus HypF is involved in regulation of hydrogenase synthesis through the HupUV proteins.
The photosynthetic bacterium Rhodobacter capsulatus contains a membrane-bound [NiFe]hydrogenase encoded by the hupSL genes. We show in this study that hypF mutants are devoid of hydrogenase activity and lack the HupL protein. We also observed that, in contrast to the wild-type strain B10, transcription of the hupSL genes was not stimulated by H2 in the hypF mutants RS13 and BSE19. Complementation of the hypF mutants with the plasmid borne hypF gene restored hydrogenase activity to wild-type levels and inducibility by H2. The R. capsulatus hupU and hupV gene products share significant similarities with the small (HupS) and the large (HupL) hydrogenase subunits, respectively. Active HupUV proteins can catalyze the hydrogen-deuterium exchange reaction. In whole cells, this H-D exchange is distinguishable from the H-D exchange catalyzed by the membrane-bound HupSL proteins by its insensitivity to O2 and to acetylene. By measuring the formation of H2 and HD in exchange with D2 uptake, we demonstrated that the hypF mutants have no active HupUV nor HupSL proteins. H-D exchange activity, of both HupUV and HupSL, was restored by hypF gene complementation. These data indicate that the HypF protein participates not only in the maturation of HupSL, but also in the maturation of the HupUV proteins and that the latter are involved in the cellular response to H2.
Colbeau
A
A
CEA/Grenoble, Laboratoire de Biochimie et Biophysique des Systèmes Intégrés (UMR CEA/CNRS no. 314), Département de Biologie Moléculaire et Structurale, France. Annette@jandri.ceng.cea.fr
Elsen
S
S
Tomiyama
M
M
Zorin
N A
NA
Dimon
B
B
Vignais
P M
PM
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D005816:Genetic Complementation Test; D006859:Hydrogen / Q000378:metabolism; D006864:Hydrogenase / Q000096:biosynthesis*; D009154:Mutation; D010088:Oxidoreductases; D010641:Phenotype; D012097:Repressor Proteins / Q000378:metabolism*; D016354:Rhodobacter capsulatus / Q000378:metabolism*
D001426:Bacterial Proteins; C101649:HupU protein, Rhodobacter capsulatus; C101650:HupV protein, Rhodobacter capsulatus; C108878:HypF protein, Bacteria; D012097:Repressor Proteins; D006859:Hydrogen; D010088:Oxidoreductases; C091789:HupL protein, Anabaena; D006864:Hydrogenase