Structural similarities and differences in H‐<scp>NS</scp> family proteins revealed by the N‐terminal structure of TurB in <i>Pseudomonas putida </i><scp>KT</scp>2440

Suzuki-Minakuchi, Chiho; Kawazuma, Kohei; Matsuzawa, Jun; Vasileva, Delyana; Fujimoto, Zui; Terada, Tohru; Okada, Kazunori; Nojiri, Hideaki

doi:10.1002/1873-3468.12425

Cited by 11 publications

(12 citation statements)

References 46 publications

(81 reference statements)

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“…1b; S1). The crystal structure of the TurB N-terminal domain (1- 61) has revealed that site 1 is formed by a “coiled-coil motif” between the N-terminal helices α1 (corresponding to H-NS α3) of two monomers 39 (fig. 1b).…”

Section: Resultsmentioning

confidence: 99%

“…1a,b). This site is also formed by hydrophobic interactions between two α-helices (α4 and α2 for the long and short members, respectively) and stabilised by salt bridges 31, 39 .…”

Section: Resultsmentioning

confidence: 99%

“…The full-length MvaT dimer was constructed by combining a homology model of the dimerization domain (residues 1-78) and the NMR structure of the DNA binding domain (PDB ID: 2MXE) 40 (residues 79-124), using Modeller version 9.15. The homology model for the dimerization domain was obtained by aligning residues 1-78 of MvaT to the crystal structure of the N-terminal domain of P. putida TurB (PDB ID: 5B52) 39 .…”

Section: Methodsmentioning

confidence: 99%

“…The upper panel depicts a schematic representation of the short H-NS oligomer. In the middle panel is the TurB N-terminal domain crystal structure (1-64) (PDB ID: 5B52) 39 . Lower panel is the MvaT protomer structural homology model (see M&M).…”

Section: Introductionmentioning

confidence: 99%

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Structural basis for osmotic regulation of the DNA binding properties of H-NS proteins

Qin

Bdira

Sterckx

et al. 2019

Preprint

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H-NS proteins act as osmotic sensors translating changes in osmolarity into altered DNA binding properties, thus, regulating enterobacterial genome organization and genes transcription. The molecular mechanism underlying the switching process and its conservation among H-NS family members remains elusive.Here, we focus on the H-NS family protein MvaT from P. aeruginosa and demonstrate experimentally that its protomer exists in two different conformations, corresponding to two different functional states. In the half-opened state (dominant at low salt) the protein forms filaments along DNA, in the fully opened state (dominant at high salt) the protein bridges DNA. This switching is a direct effect of ionic strengths on electrostatic interactions between the appositively charged DNA binding and N-terminal domains of MvaT. The asymmetric charge distribution and intramolecular interactions are conserved among the H-NS family of proteins. Therefore, our study establishes a general paradigm for the molecular mechanistic basis of the osmosensitivity of H-NS proteins.

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Section: Resultsmentioning

confidence: 99%

“…1a,b). This site is also formed by hydrophobic interactions between two α-helices (α4 and α2 for the long and short members, respectively) and stabilised by salt bridges 31, 39 .…”

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Structural basis for osmotic regulation of the DNA binding properties of H-NS proteins

Qin

Bdira

Sterckx

et al. 2019

Preprint

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“…Recently, preference of target DNA sequences of MvaT in P. aeruginosa [25] and the structure of dimerization/oligomerization domain of TurB [33] were reported. We are now performing kinetic studies on the DNA-protein and protein-protein binding properties of TurA, TurB, and Pmr.…”

Section: Discussionmentioning

confidence: 99%

Growth phase-dependent expression profiles of three vital H-NS family proteins encoded on the chromosome of Pseudomonas putida KT2440 and on the pCAR1 plasmid

et al. 2017

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BackgroundH-NS family proteins are nucleoid-associated proteins that form oligomers on DNA and function as global regulators. They are found in both bacterial chromosomes and plasmids, and were suggested to be candidate effectors of the interaction between them. TurA and TurB are the predominantly expressed H-NS family proteins encoded on the chromosome of Pseudomonas putida KT2440, while Pmr is encoded on the carbazole-degradative incompatibility group P-7 plasmid pCAR1. Previous transcriptome analyses suggested that they function cooperatively, but play different roles in the global transcriptional network. In addition to differences in protein interaction and DNA-binding functions, cell expression levels are important in clarifying the detailed underlying mechanisms. Here, we determined the precise protein amounts of TurA, TurB, and Pmr in KT2440 in the presence and absence of pCAR1.ResultsThe intracellular amounts of TurA and TurB in KT2440 and KT2440(pCAR1) were determined by quantitative western blot analysis using specific antibodies. The amount of TurA decreased from the log phase (~80,000 monomers per cell) to the stationary phase (~20,000 monomers per cell), while TurB was only detectable upon entry into the stationary phase (maximum 6000 monomers per cell). Protein amounts were not affected by pCAR1 carriage. KT2440(pCAR1pmrHis), where histidine-tagged Pmr is expressed under its original promotor, was used to determine the intracellular amount of Pmr, which was constant (~30,000 monomers per cell) during cell growth. Quantitative reverse transcription PCR demonstrated that the transcriptional levels of turA and turB were consistent with protein expression, though the transcriptional and translational profiles of Pmr differed.ConclusionThe amount of TurB increases as TurA decreases, and the amount of Pmr does not affect the amounts of TurA and TurB. This is consistent with our previous observation that TurA and TurB play complementary roles, whereas Pmr works relatively independently. This study provides insight into the molecular mechanisms underlying reconstitution of the transcriptional network in KT2440 by pCAR1 carriage.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-017-1091-6) contains supplementary material, which is available to authorized users.

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Xenogeneic Silencing and Horizontal Gene Transfer

Suzuki-Minakuchi

Navarre

2019

DNA Traffic in the Environment

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Structural similarities and differences in H‐NS family proteins revealed by the N‐terminal structure of TurB in Pseudomonas putida KT2440

Cited by 11 publications

References 46 publications

Structural basis for osmotic regulation of the DNA binding properties of H-NS proteins

Structural basis for osmotic regulation of the DNA binding properties of H-NS proteins

Growth phase-dependent expression profiles of three vital H-NS family proteins encoded on the chromosome of Pseudomonas putida KT2440 and on the pCAR1 plasmid

Xenogeneic Silencing and Horizontal Gene Transfer

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