title: Roles of AtpI and two YidC-type proteins from alkaliphilic Bacillus pseudofirmus OF4 in ATP synthase assembly and nonfermentative growth.
abstract: AtpI, a membrane protein encoded by many bacterial atp operons, is reported to be necessary for c-ring oligomer formation during assembly of some ATP synthase complexes. We investigated chaperone functions of AtpI and compared them to those of AtpZ, a protein encoded by a gene upstream of atpI that has a role in magnesium acquisition at near-neutral pH, and of SpoIIIJ and YqjG, two YidC/OxaI/Alb3 family proteins, in alkaliphilic Bacillus pseudofirmus OF4. A strain with a chromosomal deletion of atpI grew nonfermentatively, and its purified ATP synthase had a c-ring of normal size, indicating that AtpI is not absolutely required for ATP synthase function. However, deletion of atpI, but not atpZ, led to reduced stability of the ATP synthase rotor, reduced membrane association of the F(1) domain, reduced ATPase activity, and modestly reduced nonfermentative growth on malate at both pH 7.5 and 10.5. Both spoIIIJ and yqjG, but not atpI or atpZ, complemented a YidC-depleted Escherichia coli strain. Consistent with such overlapping functions, single deletions of spoIIIJ or yqjG in the alkaliphile did not affect membrane ATP synthase levels or activities, but functional specialization was indicated by YqjG and SpoIIIJ showing respectively greater roles in malate growth at pH 7.5 and 10.5. Expression of yqjG was elevated at pH 7.5 relative to that at pH 10.5 and in ΔspoIIIJ strains, but it was lower than constitutive spoIIIJ expression. Deletion of atpZ caused the largest increase among the mutants in magnesium concentrations needed for pH 7.5 growth. The basis for this phenotype is not yet resolved.
authors.affiliation: Department of Pharmacology and Systems Therapeutics, Mount Sinai School of Medicine, New York, NY, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Liu
authors.forename: David B
authors.initials: DB
authors.lastname: Hicks
authors.forename: Terry A
authors.initials: TA
authors.lastname: Krulwich
mesh_terms: D025181:ATP Synthetase Complexes / Q000378:metabolism*; D001407:Bacillus / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D017353:Gene Deletion; D005816:Genetic Complementation Test; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000378:metabolism; D008293:Malates / Q000378:metabolism; D018832:Molecular Chaperones / Q000378:metabolism*; D055503:Protein Multimerization
chemical_list: D001426:Bacterial Proteins; D008293:Malates; D018832:Molecular Chaperones; C030298:malic acid; D025181:ATP Synthetase Complexes; D008274:Magnesium