DegP protease is essential for tolerance to salt stress in the plant growth-promoting bacterium Gluconacetobacter diazotrophicus PAL5

Leandro, Mariana Ramos; Vespoli, Luciano de Souza; Andrade, Leandro Fernandes; Soares, Fabiano Silva; Boechat, Ana Laura; Pimentel, Vivian Ribeiro; Moreira, Júlia Rosa; Passamani, Lucas Zanchetta; Silveira, Vanildo; Filho, Gonçalo Apolinário de Souza

doi:10.1016/j.micres.2020.126654

Cited by 17 publications

(18 citation statements)

References 49 publications

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“…6 A, Bottom). These findings are consistent with DegP's known role as a temperature-sensitive (6,47,48) and stress-protective protease-chaperone (12,13,21).…”

Section: Discussionsupporting

confidence: 86%

“…1 B, Right) (17,22), suggesting that PDZ1:PDZ2' contacts might be critical to the formation of oligomers in the absence of substrate as well. We initially characterized the interactions between isolated PDZ domains by titrating uniformly (U)-15 N, 13 C PDZ1 (100 μM) with unlabeled PDZ2 (0 to 5 mM). Amide backbone and methyl sidechain chemical shift perturbations (CSPs), defined as differences in peak positions in the absence or presence of 5 mM PDZ2, are plotted in Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Each DegP trimer has a molecular mass of ∼140 kDa, and multiple copies of the trimer associate to form larger particles. We have therefore prepared highly deuterated protein, with 13 CH 3 labeling at Ile-δ, Leu-δ, Val-γ, and Met-e positions, where only one of the pair of isopropyl methyls of Leu and Val was 13 C-1 H heteronuclear multiple quantum coherence (HMQC) spectra that exploit the methyl-TROSY principle (39,40). We chose a high temperature (50 °C) and [NaCl] = 200 mM for obtaining the DegP concentration series, as these conditions favor path B assembly (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…1A). Highlighting its critical role in protecting bacterial function, DegP is required for growth in response to cellular stressors, such as heat (6,8), as well as oxidative (11,12) and osmotic shock (13), where it is thought to proteolyze misfolded or damaged client proteins (11,14). DegP's chaperone activities, in contrast, include the sequestration and trafficking of outer-membrane proteins (OMPs) and virulence factors through the periplasm to the outer membrane (15)(16)(17).…”

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confidence: 99%

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Competing stress-dependent oligomerization pathways regulate self-assembly of the periplasmic protease-chaperone DegP

Harkness

Toyama

Ripstein

et al. 2021

Proc. Natl. Acad. Sci. U.S.A.

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DegP is an oligomeric protein with dual protease and chaperone activity that regulates protein homeostasis and virulence factor trafficking in the periplasm of gram-negative bacteria. A number of oligomeric architectures adopted by DegP are thought to facilitate its function. For example, DegP can form a “resting” hexamer when not engaged to substrates, mitigating undesired proteolysis of cellular proteins. When bound to substrate proteins or lipid membranes, DegP has been shown to populate a variety of cage- or bowl-like oligomeric states that have increased proteolytic activity. Though a number of DegP’s substrate-engaged structures have been robustly characterized, detailed mechanistic information underpinning its remarkable oligomeric plasticity and the corresponding interplay between these dynamics and biological function has remained elusive. Here, we have used a combination of hydrodynamics and NMR spectroscopy methodologies in combination with cryogenic electron microscopy to shed light on the apo-DegP self-assembly mechanism. We find that, in the absence of bound substrates, DegP populates an ensemble of oligomeric states, mediated by self-assembly of trimers, that are distinct from those observed in the presence of substrate. The oligomeric distribution is sensitive to solution ionic strength and temperature and is shifted toward larger oligomeric assemblies under physiological conditions. Substrate proteins may guide DegP toward canonical cage-like structures by binding to these preorganized oligomers, leading to changes in conformation. The properties of DegP self-assembly identified here suggest that apo-DegP can rapidly shift its oligomeric distribution in order to respond to a variety of biological insults.

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“…6 A, Bottom). These findings are consistent with DegP's known role as a temperature-sensitive (6,47,48) and stress-protective protease-chaperone (12,13,21).…”

Section: Discussionsupporting

confidence: 86%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

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Competing stress-dependent oligomerization pathways regulate self-assembly of the periplasmic protease-chaperone DegP

Harkness

Toyama

Ripstein

et al. 2021

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…DegP assemblies can be thought of as highly dynamic and adaptive cages that are governed by a complex, client-dependent free energy landscape, where cage assemblies redistribute according to the number of bound substrate copies, client sizes, and solution conditions such as temperature. In vivo , DegP cage distributions are most likely constantly in flux within the bacterial periplasm in response to oscillations in misfolded client levels that are the result of a variety of cellular stressors including heat (5), oxidative (11), and osmotic shock perturbations (12). An understanding of the complexities of DegP’s free energy landscape is an important first step in the design of molecules to regulate its function and, potentially, to mitigate the virulence of classes of bacterial pathogens.…”

Section: Discussionmentioning

confidence: 99%

Flexible client-dependent cages in the assembly landscape of the periplasmic protease-chaperone DegP

Harkness

Ripstein

Trani

et al. 2022

Preprint

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The periplasmic protein DegP, that is implicated in virulence factor transport leading to pathogenicity, is a bi-functional protease and chaperone that maintains protein homeostasis in gram-negative bacteria. To perform these functions, DegP captures clients inside cage-like structures, which we have recently shown to form through the reorganization of high-order preformed apo-oligomers, consisting of trimeric building blocks, that are structurally distinct from client-bound cages. Our previous studies suggested that these apo oligomers may allow DegP to encapsulate clients of various sizes under protein folding stresses by forming cage ensembles that can include extremely large cage particles. To explore the relation between cage and substrate sizes, we engineered a series of DegP clients of increasing hydrodynamic radii and analyzed their influence on DegP cage formation. We used dynamic light scattering and cryogenic electron microscopy to characterize the hydrodynamic properties and structures of the DegP cages that are adopted in response to each client. We present a series of flexible cage structures including novel 30mer and 60mer particles. Key interactions between DegP trimers and the bound clients that stabilize the cage assemblies and prime the clients for catalysis are revealed. We also provide evidence that DegP can form cages which approach subcellular organelles in terms of size.

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Thermal and salt stress effects on the survival of plant growth‐promoting bacteria Azospirillum brasilense in inoculants for maize cultivation

da Cunha,

Pedrolo,

Arisi

2024

J Sci Food Agric

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BackgroundThe plant growth‐promoting bacteria (PGPB) Azospirillum brasilense is widely used as an inoculant for important grass crops, providing numerous benefits to the plants. However, one limitation to develop viable commercial inoculants is the control of PGPB survival, requiring strategies that guarantee their survival during handling and field application. The application of sublethal stress appears to be a promising strategy to increase bacterial cells tolerance to adverse environmental conditions since previous stress induces the activation of physiological protection in bacterial cell. In this work, we evaluated the effects of thermal and salt stresses on the survival of inoculant containing A. brasilense Ab‐V5 and Ab‐V6 strains and we monitored A. brasilense viability in inoculated maize roots after stress treatment of inoculant.ResultsThermal stress application (> 35 °C) in isolated cultures for both strains, as well as salt stress (NaCl concentrations > 0.3 mol.L‐1), resulted in growth rate decline. The A. brasilense enumeration in maize roots obtained by PMA‐qPCR, for inoculated maize seedlings grown in vitro for 7 days, showed that there is an increased number of viable cells after the salt stress treatment, indicating that A. brasilense Ab‐V5 and Ab‐V6 strains are able to adapt to salt stress (0.3 mol.L‐1 NaCl) growth conditions.ConclusionA. brasilense Ab‐V5 and Ab‐V6 strains had potential for osmoadaptation and salt stress, resulting in increased cell survival after inoculation in maize plants.This article is protected by copyright. All rights reserved.

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DegP protease is essential for tolerance to salt stress in the plant growth-promoting bacterium Gluconacetobacter diazotrophicus PAL5

Cited by 17 publications

References 49 publications

Competing stress-dependent oligomerization pathways regulate self-assembly of the periplasmic protease-chaperone DegP

Competing stress-dependent oligomerization pathways regulate self-assembly of the periplasmic protease-chaperone DegP

Flexible client-dependent cages in the assembly landscape of the periplasmic protease-chaperone DegP

Thermal and salt stress effects on the survival of plant growth‐promoting bacteria Azospirillum brasilense in inoculants for maize cultivation

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