Solution structure and phospho-PmrA recognition mode of PmrD from Klebsiella pneumoniae

Luo, Shih-Chi; Lou, Yuan‐Chao; Cheng, Hairong; Pan, Yun Ru; Peng, Hwei-Ling; Chen, Chinpan

doi:10.1016/j.jsb.2010.06.007

Cited by 11 publications

(14 citation statements)

References 35 publications

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“…With MTSL attached to D56C of PmrA N , three peaks corresponding to PmrD residues Gly-24 -Ala-25 and the side chain amide peak of Trp-3 were severely attenuated (data not shown). These data suggest that the interaction brings the binding surface of PmrD (the open ␤-barrel area) (18) and the active site of PmrA N close to each other. In conclusion, the observed intermolecular PRE data are generally consistent with the HADDOCK-derived complex structure (Fig.…”

Section: Pre Effect Of Site-directed Spin Labeling On Pmrdc54 Andmentioning

confidence: 79%

“…The starting structures for docking were an x-ray crystal structure of PmrA N and the 10 lowest energy structures of PmrD, which were refined with the constraints measured from the chemical shift perturbations (18). From NMR titration data, different sets of ambiguous interaction restraints were used to generate the complex model.…”

Section: Cloning Expression and Purification Of The Recombinantmentioning

confidence: 99%

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Structural Basis of a Physical Blockage Mechanism for the Interaction of Response Regulator PmrA with Connector Protein PmrD from Klebsiella pneumoniae

Luo

Lou

Rajasekaran

et al. 2013

Journal of Biological Chemistry

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Background: PmrD binds to phospho-PmrA and sustains its phosphorylation state. Results: Phospho-PmrA interacts with PmrD via several specific intermolecular interactions. Conclusion: A steric inhibition mechanism was proposed for protecting phospho-PmrA against dephosphorylation. Significance: This work provides novel data revealing how a connector protein protects an activated response regulator.

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Section: Pre Effect Of Site-directed Spin Labeling On Pmrdc54 Andmentioning

confidence: 79%

Section: Cloning Expression and Purification Of The Recombinantmentioning

confidence: 99%

Structural Basis of a Physical Blockage Mechanism for the Interaction of Response Regulator PmrA with Connector Protein PmrD from Klebsiella pneumoniae

Luo

Lou

Rajasekaran

et al. 2013

Journal of Biological Chemistry

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“…Due to its limited structural flexibility, we expect that no phosphorylation occurred. In addition, some RRs have been reported to form a homodimer after phosphorylation [ 24 , 48 ]. We did not observe the ETR1-RD to form a dimer under any conditions, even in the presence of Mg 2+ and/or the phosphoryl analog, BeF 3 - .…”

Section: Discussionmentioning

confidence: 99%

NMR Study Reveals the Receiver Domain of Arabidopsis ETHYLENE RESPONSE1 Ethylene Receptor as an Atypical Type Response Regulator

et al. 2016

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The gaseous plant hormone ethylene, recognized by plant ethylene receptors, plays a pivotal role in various aspects of plant growth and development. ETHYLENE RESPONSE1 (ETR1) is an ethylene receptor isolated from Arabidopsis and has a structure characteristic of prokaryotic two-component histidine kinase (HK) and receiver domain (RD), where the RD structurally resembles bacteria response regulators (RRs). The ETR1 HK domain has autophosphorylation activity, and little is known if the HK can transfer the phosphoryl group to the RD for receptor signaling. Unveiling the correlation of the receptor structure and phosphorylation status would advance the studies towards the underlying mechanisms of ETR1 receptor signaling. In this study, using the nuclear magnetic resonance technique, our data suggested that the ETR1-RD is monomeric in solution and the rigid structure of the RD prevents the conserved aspartate residue phosphorylation. Comparing the backbone dynamics with other RRs, we propose that backbone flexibility is critical to the RR phosphorylation. Besides the limited flexibility, ETR1-RD has a unique γ loop conformation of opposite orientation, which makes ETR1-RD unfavorable for phosphorylation. These two features explain why ETR1-RD cannot be phosphorylated and is classified as an atypical type RR. As a control, phosphorylation of the ETR1-RD was also impaired when the sequence was swapped to the fragment of the bacterial typical type RR, CheY. Here, we suggest a molecule insight that the ETR1-RD already exists as an active formation and executes its function through binding with the downstream factors without phosphorylation.

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“…Previously, we determined the solution structure of K. pneumoniae PmrD and the X-ray structure of K. pneumoniae PmrA N activated with the phosphoryl analog beryllofluoride (BeF 3 - ) and characterized their interactions by NMR and several other biophysical methods (26,27). In this study, we focused on the C-terminal DNA-binding domain.…”

Section: Introductionmentioning

confidence: 99%

Solution structure and tandem DNA recognition of the C-terminal effector domain of PmrA from Klebsiella pneumoniae

Lou

Wang

Rajasekaran

et al. 2013

Nucleic Acids Research

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Klebsiella pneumoniae PmrA is a polymyxin-resistance-associated response regulator. The C-terminal effector/DNA-binding domain of PmrA (PmrAC) recognizes tandem imperfect repeat sequences on the promoters of genes to induce antimicrobial peptide resistance after phosphorylation and dimerization of its N-terminal receiver domain (PmrAN). However, structural information concerning how phosphorylation of the response regulator enhances DNA recognition remains elusive. To gain insights, we determined the nuclear magnetic resonance solution structure of PmrAC and characterized the interactions between PmrAC or BeF3−-activated full-length PmrA (PmrAF) and two DNA sequences from the pbgP promoter of K. pneumoniae. We showed that PmrAC binds to the PmrA box, which was verified to contain two half-sites, 5′-CTTAAT-3′ and 5′-CCTAAG-3′, in a head-to-tail fashion with much stronger affinity to the first than the second site without cooperativity. The structural basis for the PmrAC–DNA complex was investigated using HADDOCK docking and confirmed by paramagnetic relaxation enhancement. Unlike PmrAC, PmrAF recognizes the two sites simultaneously and specifically. In the PmrAF–DNA complex, PmrAN may maintain an activated homodimeric conformation analogous to that in the free form and the interactions between two PmrAC molecules aid in bending and binding of the DNA duplex for transcription activation.

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Solution structure and phospho-PmrA recognition mode of PmrD from Klebsiella pneumoniae

Cited by 11 publications

References 35 publications

Structural Basis of a Physical Blockage Mechanism for the Interaction of Response Regulator PmrA with Connector Protein PmrD from Klebsiella pneumoniae

Structural Basis of a Physical Blockage Mechanism for the Interaction of Response Regulator PmrA with Connector Protein PmrD from Klebsiella pneumoniae

NMR Study Reveals the Receiver Domain of Arabidopsis ETHYLENE RESPONSE1 Ethylene Receptor as an Atypical Type Response Regulator

Solution structure and tandem DNA recognition of the C-terminal effector domain of PmrA from Klebsiella pneumoniae

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