The Activity and Specificity of the Outer Membrane Protein Chaperone SurA Are Modulated by a Proline Isomerase Domain

Ricci, Dante P.; Schwalm, Jaclyn; Gonzales-Cope, Michelle; Silhavy, Thomas J.

doi:10.1128/mbio.00540-13

Cited by 26 publications

(33 citation statements)

References 41 publications

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“…Indeed, a role for the SurA P1 domain in vivo was revealed only when surA10 was isolated in a background defective for BAM complex function (27). We thus suspected that a properly functioning BAM complex might mask any phenotype imposed by the deletion of the SurA parvulin domains.…”

Section: Deletion Of the Second Parvulin Domain Results In Inhibition Ofmentioning

confidence: 99%

“…It had been previously demonstrated that many of the OMP assembly factors, including chaperones and BAM complex members, function in overlapping and redundant pathways (4,14,27). Indeed, a role for the SurA P1 domain in vivo was revealed only when surA10 was isolated in a background defective for BAM complex function (27).…”

Section: Deletion Of the Second Parvulin Domain Results In Inhibition Ofmentioning

confidence: 99%

“…Gentle lysis was performed as previously described (27). Briefly, cultures were grown to an OD 600 of Ϸ0.8 in the presence of 0.2% L-arabinose, harvested by centrifugation, and resuspended in 100 l of 20 mM potassium phosphate (pH 7.2) and 150 mM NaCl.…”

Section: Methodsmentioning

confidence: 99%

“…Using the genetic framework established by Ricci et al (27), we report here additional genetic and biochemical evidence that establishes the P1 domain as a regulator of SurA function and OMP assembly. We use synthetic phenotypes to show that the P1 domain inhibits OMP assembly when regulation of SurA by the BAM complex is compromised.…”

mentioning

confidence: 99%

“…The bamA616 mutation confers OMP assembly defects that resemble a ⌬bamB mutant and also weakens the BamA-SurA interaction, suggesting that BamA is unable to modulate SurA activity in this strain. One suppressor obtained through this selection, termed surA10 (S220A), is a gain-of-function mutation in the P1 domain at the P1-core interface (27). Despite restoring OMP assembly in both a bamA616 and a ⌬bamB background, surA10 confers instability to the SurA protein in cell extracts, indicating that the P1 domain may inhibit SurA function and regulate its substrate repertoire.…”

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confidence: 99%

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The Activity of Escherichia coli Chaperone SurA Is Regulated by Conformational Changes Involving a Parvulin Domain

et al. 2016

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The periplasmic chaperone SurA is critical for the biogenesis of outer membrane proteins (OMPs) and, thus, the maintenance of membrane integrity in Escherichia coli. The activity of this modular chaperone has been attributed to a core chaperone module, with only minor importance assigned to the two SurA peptidyl-prolyl isomerase (PPIase) domains. In this work, we used synthetic phenotypes and covalent tethering to demonstrate that the activity of SurA is regulated by its PPIase domains and, furthermore, that its activity is correlated with the conformational state of the chaperone. When combined with mutations in the ␤-barrel assembly machine (BAM), SurA mutations resulting in deletion of the second parvulin domain (P2) inhibit OMP assembly, suggesting that P2 is involved in the regulation of SurA. The first parvulin domain (P1) potentiates this autoinhibition, as mutations that covalently tether the P1 domain to the core chaperone module severely impair OMP assembly. Furthermore, these inhibitory mutations negate the suppression of and biochemically stabilize the protein specified by a well-characterized gain-of-function mutation in P1, demonstrating that SurA cycles between distinct conformational and functional states during the OMP assembly process. IMPORTANCEThis work reveals the reversible autoinhibition of the SurA chaperone imposed by a heretofore underappreciated parvulin domain. Many ␤-barrel-associated outer membrane (OM) virulence factors, including the P-pilus and type I fimbriae, rely on SurA for proper assembly; thus, a mechanistic understanding of SurA function and inhibition may facilitate antibiotic intervention against Gram-negative pathogens, such as uropathogenic Escherichia coli, E. coli O157:H7, Shigella, and Salmonella. In addition, SurA is important for the assembly of critical OM biogenesis factors, such as the lipopolysaccharide (LPS) transport machine, suggesting that specific targeting of SurA may provide a useful means to subvert the OM barrier.T he Gram-negative bacterial outer membrane (OM) is an essential, selectively permeable barrier between the cell and its external environment. This asymmetric bilayer affords Gramnegative bacteria resistance to a wide range of antimicrobial compounds and membrane perturbants due to strong lateral interactions between lipopolysaccharides (LPS) in the outer leaflet (1, 2). In order to maintain selective permeability, the OM is replete with integral ␤-barrel outer membrane proteins (OMPs), some of which serve as pores for nutrient diffusion. The transport and assembly of these ␤-barrel proteins comprise a complicated multistep process, from their secretion through the inner membrane translocase (Sec) machinery to transport across the aqueous periplasm and assembly into the OM by the ␤-barrel assembly machine (BAM) complex (3).Prior to their assembly in the OM by BAM, these aggregationprone, insoluble ␤-barrel substrates must be maintained in a folding-competent state as they traverse the aqueous, oxidizing periplasm. This maintenance is ...

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Section: Deletion Of the Second Parvulin Domain Results In Inhibition Ofmentioning

confidence: 99%

Section: Deletion Of the Second Parvulin Domain Results In Inhibition Ofmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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The Activity of Escherichia coli Chaperone SurA Is Regulated by Conformational Changes Involving a Parvulin Domain

et al. 2016

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Analyzing the Role of Periplasmic Folding Factors in the Biogenesis of OMPs and Members of the Type V Secretion System

Bodelón

Marín

Fernández

2015

Methods in Molecular Biology

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The outer membrane (OM) of gram-negative bacteria is highly packed with OM proteins (OMPs) and the trafficking and assembly of OMPs in gram-negative bacteria is a subject of intense research. Structurally, OMPs vary in the number of β-strands and in the size and complexity of extra-membrane domains, with extreme examples being the members of the type V protein secretion system (T5SS), such as the autotransporter (AT) and intimin/invasin families of secreted proteins, in which a large extracellular "passenger" domain is linked to a β-barrel that inserts in the OM. Despite their structural and functional diversity, OMPs interact in the periplasm with a relatively small set of protein chaperones that facilitate their transport from the inner membrane (IM) to the β-barrel assembly machinery (BAM complex), preventing aggregation and assisting their folding in various aspects including disulfide bond formation. This chapter is focused on the periplasmic folding factors involved in the biogenesis of integral OMPs and members of T5SS in E. coli, which are used as a model system in this field. Background information on these periplasmic folding factors is provided along with genetic methods to generate conditional mutants that deplete these factors from E. coli and biochemical methods to analyze the folding, surface display, disulfide formation and oligomerization state of OMPs/T5SS in these mutants.

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Comparative evaluation of structure and characteristic of peptidyl-prolyl cis-trans isomerase proteins and their function in Salmonella Typhimurium stress responses and virulence

et al. 2019

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The Activity and Specificity of the Outer Membrane Protein Chaperone SurA Are Modulated by a Proline Isomerase Domain

Cited by 26 publications

References 41 publications

The Activity of Escherichia coli Chaperone SurA Is Regulated by Conformational Changes Involving a Parvulin Domain

The Activity of Escherichia coli Chaperone SurA Is Regulated by Conformational Changes Involving a Parvulin Domain

Analyzing the Role of Periplasmic Folding Factors in the Biogenesis of OMPs and Members of the Type V Secretion System

Comparative evaluation of structure and characteristic of peptidyl-prolyl cis-trans isomerase proteins and their function in Salmonella Typhimurium stress responses and virulence

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