Structural highlights
Publication Abstract from PubMed
Cell-surface signaling (CSS) in Gram-negative bacteria involves highly conserved regulatory pathways that optimize gene expression by transducing extracellular environmental signals to the cytoplasm via inner-membrane sigma regulators. The molecular details of ferric siderophore-mediated activation of the iron import machinery through a sigma regulator are unclear. Here, we present the 1.56 A resolution structure of the periplasmic complex of the C-terminal CSS domain (CCSSD) of PupR, the sigma regulator in the Pseudomonas capeferrum pseudobactin BN7/8 transport system, and the N-terminal signaling domain (NTSD) of PupB, an outer-membrane TonB-dependent transducer. The structure revealed that the CCSSD consists of two subdomains: a juxta-membrane subdomain, which has a novel all-beta fold, followed by a secretin/TonB, short N-terminal subdomain at the C-terminus of the CCSSD, a previously unobserved topological arrangement of this domain. Using affinity pull-down assays, isothermal titration calorimetry, and thermal denaturation circular dichroism spectroscopy; we show that both subdomains are required for binding the NTSD with micromolar affinity and that NTSD binding improves CCSSD stability. Our findings prompt us to present a revised model of CSS wherein the CCSSD:NTSD complex forms prior to ferric-siderophore binding. Upon siderophore binding, conformational changes in the CCSSD enable regulated intramembrane proteolysis of the sigma regulator, ultimately resulting in transcriptional regulation.
Structural basis of cell surface signaling by a conserved sigma regulator in Gram-negative bacteria.,Jensen JL, Jernberg BD, Sinha S, Colbert CL J Biol Chem. 2020 Feb 26. pii: RA119.010697. doi: 10.1074/jbc.RA119.010697. PMID:32107313[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Jensen JL, Jernberg BD, Sinha S, Colbert CL. Structural basis of cell surface signaling by a conserved sigma regulator in Gram-negative bacteria. J Biol Chem. 2020 Feb 26. pii: RA119.010697. doi: 10.1074/jbc.RA119.010697. PMID:32107313 doi:http://dx.doi.org/10.1074/jbc.RA119.010697