The WD40 Protein BamB Mediates Coupling of BAM Complexes into Assembly Precincts in the Bacterial Outer Membrane

Gunasinghe, Sachith D.; Shiota, Takuya; Stubenrauch, Christopher J.; Schulze, Keith E.; Webb, Chaille T; Fulcher, Alex J.; Dunstan, Rhys A.; Hay, Iain D.; Naderer, Thomas; Whelan, Donna R.; Bell, Toby D. M.; Elgass, Kirstin; Strugnell, Richard A.; Lithgow, Trevor

doi:10.1016/j.celrep.2018.04.093

Cited by 76 publications

(93 citation statements)

References 58 publications

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“…Consistent with this, dolP shows synthetic lethality with the gene encoding the periplasmic chaperone SurA, leading to suggestions of a role for DolP in OMP biogenesis [29][30][31] . However, we were unable to demonstrate a direct interaction between DolP and the BAM complex, and no such interaction has been seen in the extensive studies evaluating the subunit composition and multimeric states of the BAM complex [32][33][34][35] or in similar studies in N.…”

Section: Discussioncontrasting

confidence: 68%

Structure-function analyses of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localisation

Bryant

Morris

Knowles

et al. 2020

Preprint

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The Gram-negative outer membrane envelops the bacterium and functions as a permeability barrier against antibiotics, detergents and environmental stresses. Some virulence factors serve to maintain the integrity of the outer membrane, including DolP (formerly YraP) a protein of unresolved structure and function. Here we reveal DolP is a lipoprotein functionally conserved among Gram-negative bacteria and that loss of DolP increases membrane fluidity. We present the NMR solution structure for DolP, which is composed of two BON domains that form an interconnected opposing pair. The C-terminal BON domain binds to anionic phospholipids through an extensive membrane:protein interface providing evidence of subcellular localization of these phospholipids within the outer membrane. This interaction is essential for DolP function and is required for sub-cellular localization of the protein to the cell division site. The structure of DolP provides a new target for developing therapies that disrupt the integrity of the bacterial cell envelope.

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Section: Discussioncontrasting

confidence: 68%

Structure-function analyses of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localisation

Bryant

Morris

Knowles

et al. 2020

Preprint

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“…Evidence from in vivo work suggests that SurA may be involved in direct delivery of OMP substrates to BAM [19][20][21][22][23], although the molecular mechanism by which this occurs remains unresolved.…”

Section: Introductionmentioning

confidence: 99%

The Role of SurA PPIase Domains in Preventing Aggregation of the Outer-Membrane Proteins tOmpA and OmpT

Humes

Schiffrin

Calabrese

et al. 2019

Journal of Molecular Biology

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“…Although OMPs are a major component of the OM, there is scarce information about their localization and mobility (18). In E. coli, naturally abundant or overexpressed OMPs show an even distribution (19,20), whereas less abundant proteins have been shown to localize in patches (18,19,21,22) or in particular cases at the cell pole (23). These patches have been suggested to be organized in spirals (24).…”

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confidence: 99%

“…Integration of newly synthesized OMPs in the OM is accomplished by the BAM complex (27). BAM complexes have been suggested to be arranged in groups of 4 to 6 distributed across the cell, and at least one-third of the OMP patches have colocalized with a BAM complex (21,22). This organization has led to the suggestion that the BAM complexes form functional assembly precincts in the OM.…”

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confidence: 99%

Establishment of a Protein Concentration Gradient in the Outer Membrane Requires Two Diffusion-Limiting Mechanisms

et al. 2019

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OmpA-like proteins are involved in the stabilization of the outer membrane, resistance to osmotic stress, and pathogenesis. In Caulobacter crescentus, OmpA2 forms a physiologically relevant concentration gradient that forms by an uncharacterized mechanism, in which the gradient orientation depends on the position of the gene locus. This suggests that OmpA2 is synthesized and translocated to the periplasm close to the position of the gene and that the gradient forms by diffusion of the protein from this point. To further understand how the OmpA2 gradient is established, we determined the localization and mobility of the full protein and of its two structural domains. We show that OmpA2 does not diffuse and that both domains are required for gradient formation. The C-terminal domain binds tightly to the cell wall and the immobility of the full protein depends on the binding of this domain to the peptidoglycan; in contrast, the N-terminal membrane β-barrel diffuses slowly. Our results support a model in which once OmpA2 is translocated to the periplasm, the N-terminal membrane β-barrel is required for an initial fast restriction of diffusion until the position of the protein is stabilized by the binding of the C-terminal domain to the cell wall. The implications of these results on outer membrane protein diffusion and organization are discussed. IMPORTANCE Protein concentration gradients play a relevant role in the organization of the bacterial cell. The Caulobacter crescentus protein OmpA2 forms an outer membrane polar concentration gradient. To understand the molecular mechanism that determines the formation of this gradient, we characterized the mobility and localization of the full protein and of its two structural domains an integral outer membrane β-barrel and a periplasmic peptidoglycan binding domain. Each domain has a different role in the formation of the OmpA2 gradient, which occurs in two steps. We also show that the OmpA2 outer membrane β-barrel can diffuse, which is in contrast to what has been reported previously for several integral outer membrane proteins in Escherichia coli, suggesting a different organization of the outer membrane proteins.

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The WD40 Protein BamB Mediates Coupling of BAM Complexes into Assembly Precincts in the Bacterial Outer Membrane

Cited by 76 publications

References 58 publications

Structure-function analyses of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localisation

Structure-function analyses of dual-BON domain protein DolP identifies phospholipid binding as a new mechanism for protein localisation

The Role of SurA PPIase Domains in Preventing Aggregation of the Outer-Membrane Proteins tOmpA and OmpT

Establishment of a Protein Concentration Gradient in the Outer Membrane Requires Two Diffusion-Limiting Mechanisms

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