Variations in kinase and effector signaling logic in a two component signaling network DOI Creative Commons
Danielle Swingle, Lois B. Epstein, Ramisha Aymon

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 4, 2024

Abstract The general stress response (GSR) protects bacteria from a wide range of stressors. In Alphaproteobacteria , GSR activation is coordinated by HWE/HisKA2 family histidine kinases (HKs), which can exhibit non-canonical structure and function. For example, while most light-oxygen-voltage sensor-containing HKs are light activated dimers, the Rubellimicrobium thermophilum RT-HK has inverted “dark on, off” signaling logic with tunable monomer/dimer equilibrium. Here, we further investigate these atypical behaviors characterize its downstream network. Using hydrogen-deuterium exchange mass spectrometry, find that uses signal transduction mechanism similar to light-activated systems, despite logic. Mutagenesis reveals autophosphorylates in trans changes Jα helix linking sensor kinase domains affecting autophosphorylation levels. Exploring effects RT-HK, identified two genetic regions, each encoding copy central regulator PhyR. vitro measurements phosphotransfer putative PhyRs revealed signals only one, does so at an increased intensity dark, consistent reversed X-ray crystal structures both substantial shift within receiver domain suggesting basis for specificity. We probed down pathway using nuclear magnetic resonance determine single NepR homolog interacts unphosphorylated PhyRs, this interaction decoupled one This work expands our understanding transduction, revealing marked variations mechanisms previously other networks.

Language: Английский

Leveraging the histidine kinase-phosphatase duality to sculpt two-component signaling DOI Creative Commons
Stefanie Meier, Elina Multamäki, Américo Tavares Ranzani

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: June 10, 2024

Language: Английский

Citations

7
Two-component system UhpAB facilitates the pathogenicity of avian pathogenic Escherichia coli through biofilm formation and stress responses DOI
Lumin Yu, Hui Wang,

Xinglin Zhang

et al.

Avian Pathology, Journal Year: 2025, Volume and Issue: unknown, P. 1 - 12

Published: Jan. 13, 2025

Avian pathogenic Escherichia coli (APEC) is an important zoonotic pathogen that infects avian species by colonizing the gastrointestinal, respiratory, or reproductive tracts, leading to significant economic losses poultry industry worldwide and threatening food security human health. APEC has evolved two-component signal transduction system (TCS) adapt respond extracellular environmental stresses, which are produced when host invaded APEC. Here, we focus on effect of UhpAB TCS pathogenicity The results in this study showed contributed a chicken infection model. electrophoretic mobility shift assays (EMSA) confirmed specifically bound promoters fepG, ldrD, ycgV, ydeI, activated their expression, measured using real-time reverse transcription PCR (real-time RT–PCR). Furthermore, could promote biofilm formation activating expression master transcriptional regulator encoding gene csgD enhance stress tolerance protein genes uspA bhsA, thereby assisting evade immune responses inflammatory responses, increasing These findings deepen our understanding mechanism offer new perspectives for further studies prevention control infection.

Language: Английский

Citations

0
Lichtschalter zur Kontrolle bakterieller Transkription und Translation DOI Creative Commons
Marius Blackholm, Stefanie Meier, Andreas Möglich

et al.

BIOspektrum, Journal Year: 2025, Volume and Issue: 31(2), P. 154 - 157

Published: March 1, 2025

Language: Английский

Citations

0
Variations in kinase and effector signaling logic in a bacterial two component signaling network DOI Creative Commons
Danielle Swingle, Lois B. Epstein, Ramisha Aymon

et al.

Journal of Biological Chemistry, Journal Year: 2025, Volume and Issue: unknown, P. 108534 - 108534

Published: April 1, 2025

Language: Английский

Citations

0
Induction of bacterial expression at the mRNA level by light DOI Creative Commons
Américo Tavares Ranzani, K Wolburg Buchholz, Marius Blackholm

et al.

Nucleic Acids Research, Journal Year: 2024, Volume and Issue: 52(16), P. 10017 - 10028

Published: Aug. 10, 2024

Abstract Vital organismal processes, including development, differentiation and adaptation, involve altered gene expression. Although expression is frequently controlled at the transcriptional stage, various regulation mechanisms operate downstream levels. Here, we leverage photoreceptor NmPAL to optogenetically induce RNA refolding translation of bacterial mRNAs. Blue-light-triggered binding disrupts a cis-repressed mRNA state, thereby relieves obstruction initiation, upregulates Iterative probing optimization circuit, dubbed riboptoregulator, enhanced induction 30-fold. Given action level, riboptoregulator can differentially regulate individual structural genes within polycistronic operons. Moreover, it orthogonal be wed with other gene-regulatory circuits for nuanced more stringent gene-expression control. We thus advance pAurora2 circuit that combines translational increase by >1000-fold. The strategy stands upgrade numerous regulatory widely applies control in microbial biotechnology, synthetic biology materials science.

Language: Английский

Citations

2
A designed Zn2+sensor domain transmits binding information to transmembrane histidine kinases DOI Open Access
A. Katherine Hatstat, Rian Kormos,

Vee Xu

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 31, 2024

Abstract Generating stimulus-responsive, allosteric signaling de novo is a significant challenge in protein design. In natural systems like bacterial histidine kinases (HKs), signal transduction occurs when ligand binding initiates that amplified across biological membranes over long distances to induce large-scale rearrangements and phosphorylation relays. Here, we ask whether our understanding of design multi-domain, intramolecular has progressed sufficiently enable engineering HK with tunable components. We generated metal-binding sensor domains substituted them for the native domain transmembrane HK, affording chimeras transduce signals initiated from sensor. Signaling depended on designed sensor’s stability interdomain linker’s phase length. These results show usefulness elucidate biochemical mechanisms principles new systems.

Language: Английский

Citations

0
Variations in kinase and effector signaling logic in a two component signaling network DOI Creative Commons
Danielle Swingle, Lois B. Epstein, Ramisha Aymon

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 4, 2024

Abstract The general stress response (GSR) protects bacteria from a wide range of stressors. In Alphaproteobacteria , GSR activation is coordinated by HWE/HisKA2 family histidine kinases (HKs), which can exhibit non-canonical structure and function. For example, while most light-oxygen-voltage sensor-containing HKs are light activated dimers, the Rubellimicrobium thermophilum RT-HK has inverted “dark on, off” signaling logic with tunable monomer/dimer equilibrium. Here, we further investigate these atypical behaviors characterize its downstream network. Using hydrogen-deuterium exchange mass spectrometry, find that uses signal transduction mechanism similar to light-activated systems, despite logic. Mutagenesis reveals autophosphorylates in trans changes Jα helix linking sensor kinase domains affecting autophosphorylation levels. Exploring effects RT-HK, identified two genetic regions, each encoding copy central regulator PhyR. vitro measurements phosphotransfer putative PhyRs revealed signals only one, does so at an increased intensity dark, consistent reversed X-ray crystal structures both substantial shift within receiver domain suggesting basis for specificity. We probed down pathway using nuclear magnetic resonance determine single NepR homolog interacts unphosphorylated PhyRs, this interaction decoupled one This work expands our understanding transduction, revealing marked variations mechanisms previously other networks.

Language: Английский

Citations

0