title: Expression and activity of the Calvin-Benson-Bassham cycle transcriptional regulator CbbR from Acidithiobacillus ferrooxidans in Ralstonia eutropha.
abstract: Autotrophic fixation of carbon dioxide into cellular carbon occurs via several pathways but quantitatively, the Calvin-Benson-Bassham cycle is the most important. CbbR regulates the expression of the cbb genes involved in CO2 fixation via the Calvin-Benson-Bassham cycle in a number of autotrophic bacteria. A gene potentially encoding CbbR (cbbR(AF)) has been predicted in the genome of the chemolithoautotrophic, extreme acidophile Acidithiobacillus ferrooxidans. However, this microorganism is recalcitrant to genetic manipulation impeding the experimental validation of bioinformatic predictions. Two novel functional assays were devised to advance our understanding of cbbR(AF) function using the mutated facultative autotroph Ralstonia eutropha H14 ΔcbbR as a surrogate host to test gene function: (i) cbbR(AF) was expressed in R. eutropha and was able to complement ΔcbbR; and (ii) CbbR(AF) was able to regulate the in vivo activity of four A. ferrooxidans cbb operon promoters in R. eutropha. These results open up the use of R. eutropha as a surrogate host to explore cbbR(AF) activity.
authors.affiliation: Laboratorio de Biomineria, Departamento de Biotecnologia, Facultad de Ciencias del Mar y Recursos Biologicos, Universidad de Antofagasta 1240000, Chile.
authors.forename: Mario
authors.initials: M
authors.lastname: Esparza
authors.affiliation: Center for Bioinformatics and Genome Biology, Fundacion Ciencia y Vida, Santiago 7750000, Chile.
authors.forename: Eugenia
authors.initials: E
authors.lastname: Jedlicki
authors.affiliation: Centre for Ecology and Evolution in Microbial Model Systems (EEMiS), Linnaeus University, 391 82 Kalmar, Sweden.
authors.forename: Mark
authors.initials: M
authors.lastname: Dopson
authors.affiliation: Center for Bioinformatics and Genome Biology, Fundacion Ciencia y Vida, Santiago 7750000, Chile Facultad de Ciencias Biologicas, Universidad Andres Bello, Santiago 8370146, Chile dsholmes2000@yahoo.com.
authors.forename: David S
authors.initials: DS
authors.lastname: Holmes
mesh_terms: D042763:Acidithiobacillus / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D057486:Carbon Cycle; D003001:Cloning, Molecular; D020118:Cupriavidus necator / Q000235:genetics* / Q000502:physiology; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D009876:Operon; D010788:Photosynthesis / Q000235:genetics* / Q000502:physiology; D011401:Promoter Regions, Genetic; D050436:Regulatory Elements, Transcriptional; D016415:Sequence Alignment; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D014157:Transcription Factors; C072714:cfxR protein, bacteria
keywords: Acidithiobacillus ferrooxidans; CO2 fixation; CbbR; Ralstonia eutropha; RubisCO; autotroph