title: First molecular characterisation of hydrogenosomes in the protozoan parasite Histomonas meleagridis.
abstract: Histomonas meleagridis is a trichomonad species that undergoes a flagellate-to-amoeba transformation during tissue invasion and causes a serious disease in gallinaceous birds (blackhead disease or histomoniasis). Living in the avian cecum, the flagellated form can be grown in vitro in the presence of an ill-defined bacterial flora. Its cytoplasm harbours numerous spherical bodies which structurally resemble hydrogenosomes. To test whether these organelles may be involved in anaerobic metabolism, we undertook the identification of H. meleagridis genes encoding some potentially conserved hydrogenosomal enzymes. The strategy was based on several PCR amplification steps using primers designed from available sequences of the phylogenetically-related human parasite Trichomonas vaginalis. We first obtained a C-terminal sequence of an iron-hydrogenase homologue (Hm_HYD) with typical active site signatures (H-cluster domain). Immunoelectron microscopy with anti-Hm_HYD polyclonal antibodies showed specific gold labelling of electron-dense organelles, thus confirming their hydrogenosomal nature. The whole genes encoding a malic enzyme (Hm_ME) and the alpha-subunit of a succinyl coenzyme A synthetase (Hm_alpha-SCS) were then identified. Short N-terminal presequences for hydrogenosomal targeting were predicted in both proteins. Anti-Hm_ME and anti-Hm_alpha-SCS antisera provided immunofluorescence staining patterns of H. meleagridis cytoplasmic granules similar to those observed with anti-Hm_HYD antiserum or mAb F5.2 known to react with T. vaginalis hydrogenosomes. Hm_ME, Hm_alpha-SCS and Hm_HYD were also detected as reactive bands on immunoblots of proteins from purified hydrogenosomes. Interestingly, anti-Hm_alpha-SCS staining of the cell surface in non-permeabilised parasites suggests a supplementary role for SCS in cytoadherence, as previously demonstrated in T. vaginalis.
authors: Equipe Parasitologie Moléculaire et Cellulaire, LBP, UMR CNRS 6023, Université Blaise Pascal, 24 Avenue des Landais, 63177 Aubière Cedex, France.
authors: Muriel
authors: M
authors: Mazet
authors: Marie
authors: M
authors: Diogon
authors: John F
authors: JF
authors: Alderete
authors: Christian P
authors: CP
authors: Vivarès
authors: Frédéric
authors: F
authors: Delbac
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002448:Cell Adhesion; D005455:Fluorescent Antibody Technique; D017125:Genes, Protozoan; D006859:Hydrogen / Q000378:metabolism*; D006864:Hydrogenase / Q000235:genetics; D007150:Immunohistochemistry; D007506:Iron-Sulfur Proteins / Q000235:genetics; D008291:Malate Dehydrogenase / Q000235:genetics; D008969:Molecular Sequence Data; D015388:Organelles / Q000201:enzymology / Q000235:genetics*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013389:Succinate-CoA Ligases / Q000235:genetics; D014244:Trichomonas / Q000201:enzymology / Q000235:genetics*; D014246:Trichomonas vaginalis / Q000201:enzymology / Q000235:genetics
chemical_list: D007506:Iron-Sulfur Proteins; D006859:Hydrogen; D008291:Malate Dehydrogenase; C118830:iron hydrogenase; D006864:Hydrogenase; D013389:Succinate-CoA Ligases