scholarly journals PATAN-domain response regulators interact with the Type IV pilus motor to control phototactic orientation in the cyanobacterium Synechocystis sp. PCC 6803

2021 ◽  
Author(s):  
Yu Han ◽  
Annik Jakob ◽  
Sophia Engel ◽  
Annegret Wilde ◽  
Nils Schuergers
Keyword(s):  
Response Regulator ◽  
Cellular Protein ◽  
Temporal Organization ◽  
Protein Distribution ◽  
Response Regulators ◽  
Type Iv Pilus ◽  
Signaling Systems ◽  
Type Iv ◽  
Chemosensory Systems ◽  
Output Domain

Many prokaryotes show complex behaviors that require the intricate spatial and temporal organization of cellular protein machineries, leading to asymmetrical protein distribution and cell polarity. One such behavior is cyanobacterial phototaxis which relies on the dynamic localization of the Type IV pilus motor proteins in response to light. In the cyanobacterium Synechocystis, various signaling systems encompassing chemotaxis-related CheY- and PatA-like response regulators are critical players in switching between positive and negative phototaxis depending on the light intensity and wavelength. In this study, we show that PatA-type regulators evolved from chemosensory systems. Using fluorescence microscopy and yeast-two-hybrid analysis, we demonstrate that they localize to the inner membrane, where they interact with the N-terminal cytoplasmic domain of PilC and the pilus assembly ATPase PilB1. By separately expressing the subdomains of the response regulator PixE, we confirm that only the N-terminal PATAN domain interacts with PilB1, localizes to the membrane, and is sufficient to reverse phototactic orientation. These experiments established that the PATAN domain is the principal output domain of PatA-type regulators which we presume to modulate pilus extension by binding to the pilus motor components.

scholarly journals Structural basis of response regulator function

2019 ◽  
Author(s):  
Rong Gao ◽  
Sophie Bouillet ◽  
Ann Stock
Keyword(s):  
Modular Design ◽  
Response Regulator ◽  
Structural Features ◽  
Structural Basis ◽  
Adaptive Responses ◽  
Response Regulators ◽  
Signaling Systems ◽  
Regulatory Strategies ◽  
Two Component Systems ◽  
Functional Features

Response regulators function as the output components of two-component systems, which couple the sensing of environmental stimuli to adaptive responses. Response regulators typically contain conserved receiver (REC) domains that function as phosphorylation-regulated switches to control the activities of effector domains that elicit output responses. This modular design is extremely versatile, enabling different regulatory strategies tuned to the needs of individual signaling systems. This review summarizes functional features that underlie response regulator function. An abundance of atomic resolution structures and complementary biochemical data have defined the mechanisms for response regulator enzymatic activities, revealed trends in regulatory strategies utilized by response regulators of different subfamilies and provided insights into interactions of response regulators with their cognate histidine kinases. Among the hundreds of thousands of response regulators identified, variations abound. This article provides a framework for understanding structural features that enable function of canonical response regulators and a basis for distinguishing non-canonical configurations.

scholarly journals Structural basis of response regulator function

2019 ◽  
Author(s):  
Rong Gao ◽  
Sophie Bouillet ◽  
Ann Stock
Keyword(s):  
Modular Design ◽  
Response Regulator ◽  
Structural Features ◽  
Structural Basis ◽  
Adaptive Responses ◽  
Response Regulators ◽  
Signaling Systems ◽  
Regulatory Strategies ◽  
Two Component Systems ◽  
Functional Features

Response regulators function as the output components of two-component systems, which couple the sensing of environmental stimuli to adaptive responses. Response regulators typically contain conserved receiver (REC) domains that function as phosphorylation-regulated switches to control the activities of effector domains that elicit output responses. This modular design is extremely versatile, enabling different regulatory strategies tuned to the needs of individual signaling systems. This review summarizes functional features that underlie response regulator function. An abundance of atomic resolution structures and complementary biochemical data have defined the mechanisms for response regulator enzymatic activities, revealed trends in regulatory strategies utilized by response regulators of different subfamilies and provided insights into interactions of response regulators with their cognate histidine kinases. Among the hundreds of thousands of response regulators identified, variations abound. This article provides a framework for understanding structural features that enable function of canonical response regulators and a basis for distinguishing non-canonical configurations.

scholarly journals The Orphan Response Regulator DigR Is Required for Synthesis of Extracellular Matrix Fibrils in Myxococcus xanthus

2006 ◽  
Vol 188 (12) ◽  
pp. 4384-4394 ◽  
Author(s):  
Martin Overgaard ◽  
Sigrun Wegener-Feldbrügge ◽  
Lotte Søgaard-Andersen
Keyword(s):  
Extracellular Matrix ◽  
Response Regulator ◽  
Cell Envelope ◽  
Transcriptional Profiling ◽  
Myxococcus Xanthus ◽  
Gliding Motility ◽  
Differentially Expressed ◽  
Type Iv Pilus ◽  
Receiver Domain ◽  
Type Iv

ABSTRACT In Myxococcus xanthus, two-component systems have crucial roles in regulating motility behavior and development. Here we describe an orphan response regulator, consisting of an N-terminal receiver domain and a C-terminal DNA binding domain, which is required for A and type IV pilus-dependent gliding motility. Genetic evidence suggests that phosphorylation of the conserved, phosphorylatable aspartate residue in the receiver domain is required for DigR activity. Consistent with the defect in type IV pilus-dependent motility, a digR mutant is slightly reduced in type IV pilus biosynthesis, and the composition of the extracellular matrix fibrils is abnormal, with an increased content of polysaccharides and decreased accumulation of the FibA metalloprotease. By using genome-wide transcriptional profiling, 118 genes were identified that are directly or indirectly regulated by DigR. These 118 genes include only 2, agmQ and cheY4, previously implicated in A and type IV pilus-dependent motility, respectively. In silico analyses showed that 36% of the differentially expressed genes are likely to encode exported proteins. Moreover, four genes encoding homologs of extracytoplasmic function (ECF) sigma factors, which typically control aspects of cell envelope homeostasis, are differentially expressed in a digR mutant. We suggest that the DigR response regulator has an important function in cell envelope homeostasis and that the motility defects in a digR mutant are instigated by the abnormal cell envelope and abnormal expression of agmQ and cheY4.

scholarly journals Genetic Evidence that the α5 Helix of the Receiver Domain of PhoB Is Involved in Interdomain Interactions

2001 ◽  
Vol 183 (7) ◽  
pp. 2204-2211 ◽  
Author(s):  
Mindy P. Allen ◽  
Kimberly B. Zumbrennen ◽  
William R. McCleary
Keyword(s):  
Response Regulator ◽  
Response Regulators ◽  
Receiver Domain ◽  
Intracellular Signals ◽  
Regulator Protein ◽  
Phosphorylation Cascade ◽  
Α Helix ◽  
Interdomain Interactions ◽  
Response Regulator Protein ◽  
Output Domain

ABSTRACT Two-component signaling proteins are involved in transducing environmental stimuli into intracellular signals. Information is transmitted through a phosphorylation cascade that consists of a histidine protein kinase and a response regulator protein. Generally, response regulators are made up of a receiver domain and an output domain. Phosphorylation of the receiver domain modulates the activity of the output domain. The mechanisms by which receiver domains control the activities of their respective output domains are unknown. To address this question for the PhoB protein from Escherichia coli, we have employed two separate genetic approaches, deletion analysis and domain swapping. In-frame deletions were generated within the phoB gene, and the phenotypes of the mutants were analyzed. The output domain, by itself, retained significant ability to activate transcription of the phoA gene. However, another deletion mutant that contained the C-terminal α-helix of the receiver domain (α5) in addition to the entire output domain was unable to activate transcription of phoA. This result suggests that the α5 helix of the receiver domain interacts with and inhibits the output domain. We also constructed two chimeric proteins that join various parts of the chemotaxis response regulator, CheY, to PhoB. A chimera that joins the N-terminal ∼85% of CheY's receiver domain to the β5-α5 loop of PhoB's receiver domain displayed phosphorylation-dependent activity. The results from both sets of experiments suggest that the regulation of PhoB involves the phosphorylation-mediated modulation of inhibitory contacts between the α5 helix of its unphosphorylated receiver domain and its output domain.

scholarly journals Lysobacter PilR, the Regulator of Type IV Pilus Synthesis, Controls Antifungal Antibiotic Production via a Cyclic di-GMP Pathway

2017 ◽  
Vol 83 (7) ◽  
Author(s):  
Yuan Chen ◽  
Jing Xia ◽  
Zhenhe Su ◽  
Gaoge Xu ◽  
Mark Gomelsky ◽  
...  
Keyword(s):  
Response Regulator ◽  
Antibiotic Production ◽  
Second Messenger ◽  
Twitching Motility ◽  
Type Iv Pilus ◽  
Content Type ◽  
Lysobacter Enzymogenes ◽  
Antifungal Antibiotic ◽  
Type Iv ◽  
Two Component

ABSTRACT Lysobacter enzymogenes is a ubiquitous soil gammaproteobacterium that produces a broad-spectrum antifungal antibiotic, known as heat-stable antifungal factor (HSAF). To increase HSAF production for use against fungal crop diseases, it is important to understand how HSAF synthesis is regulated. To gain insights into transcriptional regulation of the HSAF synthesis gene cluster, we generated a library with deletion mutations in the genes predicted to encode response regulators of the two-component signaling systems in L. enzymogenes strain OH11. By quantifying HSAF production levels in the 45 constructed mutants, we identified two strains that produced significantly smaller amounts of HSAF. One of the mutations affected a gene encoding a conserved bacterial response regulator, PilR, which is commonly associated with type IV pilus synthesis. We determined that L. enzymogenes PilR regulates pilus synthesis and twitching motility via a traditional pathway, by binding to the pilA promoter and upregulating pilA expression. Regulation of HSAF production by PilR was found to be independent of pilus formation. We discovered that the pilR mutant contained significantly higher intracellular levels of the second messenger cyclic di-GMP (c-di-GMP) and that this was the inhibitory signal for HSAF production. Therefore, the type IV pilus regulator PilR in L. enzymogenes activates twitching motility while downregulating antibiotic HSAF production by increasing intracellular c-di-GMP levels. This study identifies a new role of a common pilus regulator in proteobacteria and provides guidance for increasing antifungal antibiotic production in L. enzymogenes. IMPORTANCE PilR is a widespread response regulator of the two-component system known for regulating type IV pilus synthesis in proteobacteria. Here we report that, in the soil bacterium Lysobacter enzymogenes, PilR regulates pilus synthesis and twitching motility, as expected. Unexpectedly, PilR was also found to control intracellular levels of the second messenger c-di-GMP, which in turn inhibits production of the antifungal antibiotic HSAF. The coordinated production of type IV pili and antifungal antibiotics has not been observed previously.

Structural Basis of Response Regulator Function

2019 ◽  
Vol 73 (1) ◽  
pp. 175-197 ◽  
Author(s):  
Rong Gao ◽  
Sophie Bouillet ◽  
Ann M. Stock
Keyword(s):  
Modular Design ◽  
Response Regulator ◽  
Structural Features ◽  
Structural Basis ◽  
Adaptive Responses ◽  
Response Regulators ◽  
Signaling Systems ◽  
Regulatory Strategies ◽  
Component Systems ◽  
Two Component Systems

Response regulators function as the output components of two-component systems, which couple the sensing of environmental stimuli to adaptive responses. Response regulators typically contain conserved receiver (REC) domains that function as phosphorylation-regulated switches to control the activities of effector domains that elicit output responses. This modular design is extremely versatile, enabling different regulatory strategies tuned to the needs of individual signaling systems. This review summarizes structural features that underlie response regulator function. An abundance of atomic resolution structures and complementary biochemical data have defined the mechanisms for response regulator enzymatic activities, revealed trends in regulatory strategies utilized by response regulators of different subfamilies, and provided insights into interactions of response regulators with their cognate histidine kinases. Among the hundreds of thousands of response regulators identified, variations abound. This article provides a framework for understanding structural features that enable function of canonical response regulators and a basis for distinguishing noncanonical configurations.

scholarly journals Pseudomonas aeruginosa AlgR Regulates Type IV Pilus Biosynthesis by Activating Transcription of the fimU-pilVWXY1Y2E Operon

2008 ◽  
Vol 190 (6) ◽  
pp. 2023-2030 ◽  
Author(s):  
Belen Belete ◽  
Haiping Lu ◽  
Daniel J. Wozniak
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Response Regulator ◽  
Transcriptional Profiling ◽  
Pcr Analysis ◽  
Twitching Motility ◽  
Type Iv Pilus ◽  
Mobility Shift ◽  
Type Iv ◽  
Type Iv Pilin ◽  
Electrophoretic Mobility Shift Assays

ABSTRACT The response regulator AlgR is required for Pseudomonas aeruginosa type IV pilus-dependent twitching motility, a flagellum-independent mode of solid surface translocation. Prior work showed that AlgR is phosphorylated at aspartate 54, and cells expressing an AlgR variant that cannot undergo phosphorylation (AlgRD54N) lack twitching motility. However, the mechanism by which AlgR controls twitching motility is not completely understood. We hypothesized that AlgR functioned by activating genes within the prepilin fimU-pilVWXY1Y2E cluster that are necessary for type IV pilin biogenesis. Reverse transcriptase PCR analysis showed that the fimU-pilVWXY1Y2E genes are cotranscribed in an operon, which is under the control of AlgR. This supports prior transcriptional profiling studies of wild-type strains and algR mutants. Moreover, expression of the fimU-pilVWXY1Y2E operon was reduced in strains expressing AlgRD54N. DNase footprinting and electrophoretic mobility shift assays demonstrate that AlgR but not AlgRD54N bound with high affinity to two sites upstream of the fimU-pilVWXY1Y2E operon. Altogether, our findings indicate that AlgR is essential for proper pilin localization and that phosphorylation of AlgR results in direct activation of the fimU-pilVWXY1Y2E operon, which is required for the assembly and export of a functional type IV pilus.

scholarly journals Structural basis of response regulator function

2019 ◽  
Author(s):  
Rong Gao ◽  
Sophie Bouillet ◽  
Ann Stock
Keyword(s):  
Modular Design ◽  
Response Regulator ◽  
Structural Features ◽  
Structural Basis ◽  
Adaptive Responses ◽  
Response Regulators ◽  
Signaling Systems ◽  
Regulatory Strategies ◽  
Two Component Systems ◽  
Functional Features

Response regulators function as the output components of two-component systems, which couple the sensing of environmental stimuli to adaptive responses. Response regulators typically contain conserved receiver (REC) domains that function as phosphorylation-regulated switches to control the activities of effector domains that elicit output responses. This modular design is extremely versatile, enabling different regulatory strategies tuned to the needs of individual signaling systems. This review summarizes functional features that underlie response regulator function. An abundance of atomic resolution structures and complementary biochemical data have defined the mechanisms for response regulator enzymatic activities, revealed trends in regulatory strategies utilized by response regulators of different subfamilies and provided insights into interactions of response regulators with their cognate histidine kinases. Among the hundreds of thousands of response regulators identified, variations abound. This article provides a framework for understanding structural features that enable function of canonical response regulators and a basis for distinguishing non-canonical configurations.

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