title: Gamma carbonic anhydrases in plant mitochondria.
abstract: Three genes from Arabidopsis thaliana with high sequence similarity to gamma carbonic anhydrase (gammaCA), a Zn containing enzyme from Methanosarcina thermophila (CAM), were identified and characterized. Evolutionary and structural analyses predict that these genes code for active forms of gammaCA. Phylogenetic analyses reveal that these Arabidopsis gene products cluster together with CAM and related sequences from alpha and gamma proteobacteria, organisms proposed as the mitochondrial endosymbiont ancestor. Indeed, in vitro and in vivo experiments indicate that these gene products are transported into the mitochondria as occurs with several mitochondrial protein genes transferred, during evolution, from the endosymbiotic bacteria to the host genome. Moreover, putative CAM orthologous genes are detected in other plants and green algae and were predicted to be imported to mitochondria. Structural modeling and sequence analysis performed in more than a hundred homologous sequences show a high conservation of functionally important active site residues. Thus, the three histidine residues involved in Zn coordination (His 81, 117 and 122), Arg 59, Asp 61, Gin 75, and Asp 76 of CAM are conserved and properly arranged in the active site cavity of the models. Two other functionally important residues (Glu 62 and Glu 84 of CAM) are lacking, but alternative amino acids that might serve to their roles are postulated. Accordingly, we propose that photosynthetic eukaryotic organisms (green algae and plants) contain gammaCAs and that these enzymes codified by nuclear genes are imported into mitochondria to accomplish their biological function.
authors.affiliation: Centro de Estudios de Investigaciones, Universidad Nacional de Quilmes, Roque Sáenz Peña 180, Bernal, Argentina.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Parisi
authors.forename: Mariano
authors.initials: M
authors.lastname: Perales
authors.forename: María Silvina
authors.initials: MS
authors.lastname: Fornasari
authors.forename: Alejandro
authors.initials: A
authors.lastname: Colaneri
authors.forename: Nahuel
authors.initials: N
authors.lastname: González-Schain
authors.forename: Diego
authors.initials: D
authors.lastname: Gómez-Casati
authors.forename: Sabrina
authors.initials: S
authors.lastname: Zimmermann
authors.forename: Axel
authors.initials: A
authors.lastname: Brennicke
authors.forename: Alejandro
authors.initials: A
authors.lastname: Araya
authors.forename: James G
authors.initials: JG
authors.lastname: Ferry
authors.forename: Julián
authors.initials: J
authors.lastname: Echave
authors.forename: Eduardo
authors.initials: E
authors.lastname: Zabaleta
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D019843:Archaeal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017020:Methanosarcina / Q000235:genetics; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010802:Phylogeny; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D029681:Arabidopsis Proteins; D019843:Archaeal Proteins; D007527:Isoenzymes; D024101:Mitochondrial Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D002256:Carbonic Anhydrases