title: In vivo roles of BamA, BamB and BamD in the biogenesis of BamA, a core protein of the β-barrel assembly machine of Escherichia coli.
abstract: Assembly of the β-barrel outer membrane proteins (OMPs) is an essential cellular process in Gram-negative bacteria and in the mitochondria and chloroplasts of eukaryotes--two organelles of bacterial origin. Central to this process is the conserved β-barrel OMP that belongs to the Omp85 superfamily. In Escherichia coli, BamA is the core β-barrel OMP and, together with four outer membrane lipoproteins, BamBCDE, constitutes the β-barrel assembly machine (BAM). In this paper, we investigated the roles of BamD, an essential lipoprotein, and BamB in BamA biogenesis. Depletion of BamD caused impairment in BamA biogenesis and cessation of cell growth. These defects of BamD depletion were partly reversed by single-amino-acid substitutions mapping within the β-barrel domain of BamA. However, in the absence of BamB, the positive effects of the β-barrel substitutions on BamA biogenesis under BamD depletion conditions were nullified. By employing a BamA protein bearing one such substitution, F474L, it was demonstrated that the mutant BamA protein could not only assemble without BamD but also facilitate the assembly of wild-type BamA expressed in trans. Based on these data, we propose a model in which the Bam lipoproteins, which are localized to the outer membrane by the BAM-independent Lol pathway, aid in the creation of new BAM complexes by serving as outer membrane receptors and folding factors for nascent BamA molecules. The newly assembled BAM holocomplex then catalyzes the assembly of substrate OMPs and BamA. These in vivo findings are corroborated by recently published in vitro data.
authors.affiliation: School of Life Sciences, Arizona State University, Tempe, AZ 85287, USA. Electronic address: rajeev.misra@asu.edu.
authors.forename: Rajeev
authors.initials: R
authors.lastname: Misra
authors.affiliation: School of Life Sciences, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Stikeleather
authors.affiliation: School of Life Sciences, Arizona State University, Tempe, AZ 85287, USA.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Gabriele
mesh_terms: D019943:Amino Acid Substitution / Q000502:physiology; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism*; D008074:Lipoproteins / Q000378:metabolism; D001678:Organelle Biogenesis; D017510:Protein Folding
chemical_list: D001425:Bacterial Outer Membrane Proteins; C503914:BamA protein, E coli; C557407:BamB protein, E coli; C513739:BamD protein, E coli; D029968:Escherichia coli Proteins; D008074:Lipoproteins
keywords: BAM complex; genetics; β-barrel outer membrane protein assembly