Literature summary for 2.7.13.3 extracted from

Chambonnier, G.; Roux, L.; Redelberger, D.; Fadel, F.; Filloux, A.; Sivaneson, M.; de Bentzmann, S.; Bordi, C.
The hybrid histidine kinase LadS forms a multicomponent signal transduction system with the GacS/GacA two-component system in Pseudomonas aeruginosa (2016), PLoS Genet., 12, e1006032.

Search for more literature for 2.7.13.3

Cloned(Commentary)

Cloned (Comment)	Organism
recombinant expression of C-terminally His-tagged forms of enzyme domains GacSD1, GacSH1D1, GacSH2, and GacSH2H->Q in Escherichia coli strain BTH101, expression of N-terminally FLAG or Strep tagged enzyme fragment constructs	Pseudomonas aeruginosa
recombinant expression of C-terminally His-tagged forms of enzyme domains LadSH1D1, LadSH1D1D->A, and LadSD1 in Escherichia coli strain BTH101, expression of N-terminally FLAG or Strep tagged enzyme fragment constructs	Pseudomonas aeruginosa

Protein Variants

Protein Variants	Comment	Organism
additional information	construction of diverse point, truncation, and deletion mutants of enzyme GacS, and generation of diverse two-hybrid constructs of enzyme GacS domains	Pseudomonas aeruginosa
additional information	construction of diverse point, truncation, and deletion mutants of enzyme LadS, and generation of diverse two-hybrid constructs of enzyme LadS domains	Pseudomonas aeruginosa

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
inner membrane	-	Pseudomonas aeruginosa	-	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Mg2+	required	Pseudomonas aeruginosa

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
ATP + protein L-histidine	Pseudomonas aeruginosa	-	ADP + protein N-phospho-L-histidine	-	?
additional information	Pseudomonas aeruginosa	A two-component system comprises a histidine kinase protein or sensor mostly inserted into the inner membrane and a cognate partner known as the response regulator. The stimulus by the periplasmic or cytoplasmic detection domain of the histidine kinase protein triggers autophosphorylation on a conserved histidine residue of the transmitter domain H1. The phosphoryl group is then transferred on a conserved aspartate residue present in the receiver or D domain of the cognate response regulator. In Pseudomonas aeruginosa, the histidine kinase requires additional domains such as a receiver domain (D1) fused to the histidine kinase. Two-component system mechanism, overview	?	-	?
additional information	Pseudomonas aeruginosa	a two-component system comprises a histidine kinase protein or sensor mostly inserted into the inner membrane and a cognate partner known as the response regulator. The stimulus by the periplasmic or cytoplasmic detection domain of the histidine kinase protein triggers autophosphorylation on a conserved histidine residue of the transmitter domain H1. The phosphoryl group is then transferred on a conserved aspartate residue present in the receiver or D domain of the cognate response regulator. In Pseudomonas aeruginosa, the histidine kinase requires additional domains such as a receiver domain (D1) fused to the histidine kinase. Two-component system mechanism, overview. GacS is an unorthodox histidine kinase with H1/D1/H2 domains. GacA is an response regulator functioning as a transcriptional regulator, which positively and exclusively controls the expression of two unique target genes encoding two small noncoding RNAs, RsmY and RsmZ	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas aeruginosa	G3XD98	-	-
Pseudomonas aeruginosa	Q9HX42	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
additional information	in vitro transphosphorylation of GacS H2 domain by LadS histidine kinase	Pseudomonas aeruginosa

Purification (Commentary)

Purification (Comment)	Organism
recombinant C-terminally His-tagged forms of enzyme domains GacSD1, GacSH1D1, GacSH2, and GacSH2H->Q from Escherichia coli strain BTH101 by nickel affinity chromatography	Pseudomonas aeruginosa
recombinant C-terminally His-tagged forms of enzyme domains LadSH1D1, LadSH1D1D->A, and LadSD1 from Escherichia coli strain BTH101 by nickel affinity chromatography	Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
ATP + protein L-histidine	-	Pseudomonas aeruginosa	ADP + protein N-phospho-L-histidine	-	?
additional information	A two-component system comprises a histidine kinase protein or sensor mostly inserted into the inner membrane and a cognate partner known as the response regulator. The stimulus by the periplasmic or cytoplasmic detection domain of the histidine kinase protein triggers autophosphorylation on a conserved histidine residue of the transmitter domain H1. The phosphoryl group is then transferred on a conserved aspartate residue present in the receiver or D domain of the cognate response regulator. In Pseudomonas aeruginosa, the histidine kinase requires additional domains such as a receiver domain (D1) fused to the histidine kinase. Two-component system mechanism, overview	Pseudomonas aeruginosa	?	-	?
additional information	a two-component system comprises a histidine kinase protein or sensor mostly inserted into the inner membrane and a cognate partner known as the response regulator. The stimulus by the periplasmic or cytoplasmic detection domain of the histidine kinase protein triggers autophosphorylation on a conserved histidine residue of the transmitter domain H1. The phosphoryl group is then transferred on a conserved aspartate residue present in the receiver or D domain of the cognate response regulator. In Pseudomonas aeruginosa, the histidine kinase requires additional domains such as a receiver domain (D1) fused to the histidine kinase. Two-component system mechanism, overview. GacS is an unorthodox histidine kinase with H1/D1/H2 domains. GacA is an response regulator functioning as a transcriptional regulator, which positively and exclusively controls the expression of two unique target genes encoding two small noncoding RNAs, RsmY and RsmZ	Pseudomonas aeruginosa	?	-	?
additional information	in vitro transphosphorylation of GacS H2 domain by LadS histidine kinase. The enzyme performs autophosphorylation using ATP	Pseudomonas aeruginosa	?	-	?

Synonyms

Synonyms	Comment	Organism
GacS	-	Pseudomonas aeruginosa
LadS	-	Pseudomonas aeruginosa
PA0928	-	Pseudomonas aeruginosa
PA3974	-	Pseudomonas aeruginosa

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
22	-	assay at room temperature	Pseudomonas aeruginosa

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.6	-	assay at	Pseudomonas aeruginosa

Cofactor

Cofactor	Comment	Organism	Structure
ATP	-	Pseudomonas aeruginosa

General Information

General Information	Comment	Organism
evolution	while the GacS/GacA two-component system is widely distributed throughout the bacterial kingdom, the molecular switch formed by the hybrid LadS, PA1611 and RetS histifine kinases is unique to the Pseudomonas species, though it can function in very different ways in phylogenetically related Pseudomonas species	Pseudomonas aeruginosa
malfunction	gacS or gacA mutations are epistasic to ladS mutation	Pseudomonas aeruginosa
physiological function	in response to environmental changes, Pseudomonas aeruginosa is able to switch from a planktonic (free swimming) to a sessile (biofilm) lifestyle. The two-component system GacS/GacA activates the production of two small non-coding RNAs, RsmY and RsmZ, but four histidine kinases, RetS, GacS, LadS and PA1611, are instrumental in this process. RetS hybrid histidine kinase blocks GacS unorthodox histidine kinase autophosphorylation through the formation of a heterodimer. PA1611 hybrid histidine kinase, which is structurally related to GacS, interacts with RetS in Pseudomonas aeruginosa in a very similar manner to GacS. LadS hybrid histidine kinase phenotypically antagonizes the function of RetS by a mechanism that has never been investigated. The four sensors are found in most Pseudomonas species but their characteristics and mode of signaling may differ from one species to another. In Pseudomonas aeruginosa, LadS controls both rsmY and rsmZ gene expression and this regulation occurs through the GacS/GacA two-component system. In contrast to RetS, LadS signals through GacS/GacA without forming heterodimers, either with GacS or with RetS. Enzyme LadS is involved in a genuine phospho relay, which requires both transmitter and receiver LadS domains. LadS signaling ultimately requires the alternative histidine-phosphotransfer domain of GacS. LadS histidine kinase forms, with the GacS/GacA two-component system, a multicomponent signal transduction system with an original phosphorelay cascade, i.e. H1LadS -> D1LadS -> H2GacS -> D2GacA. This highlights an original strategy in which a unique output, i.e. the modulation of sRNA levels, is controlled by a complex multi-sensing network to fine-tune an adapted biofilm and virulence response. H1 and D1 domain involvement of the LadS hybrid histidine kinase in the LadS signaling pathway, and involvement of the H2 domain of the GacS unorthodox histidine kinase in the LadS signaling pathway, overview	Pseudomonas aeruginosa
physiological function	in response to environmental changes, Pseudomonas aeruginosa is able to switch from a planktonic (free swimming) to a sessile (biofilm) lifestyle. The two-component system GacS/GacA activates the production of two small non-coding RNAs, RsmY and RsmZ, but four histidine kinases, RetS, GacS, LadS and PA1611, are instrumental in this process. RetS hybrid histidine kinase blocks GacS unorthodox histidine kinase autophosphorylation through the formation of a heterodimer. PA1611 hybrid histidine kinase, which is structurally related to GacS, interacts with RetS in Pseudomonas aeruginosa in a very similar manner to GacS. LadS hybrid histidine kinase phenotypically antagonizes the function of RetS. The four sensors are found in most Pseudomonas species but their characteristics and mode of signaling may differ from one species to another. In Pseudomonas aeruginosa, LadS controls both rsmY and rsmZ gene expression and this regulation occurs through the GacS/GacA two-component system. In contrast to RetS, LadS signals through GacS/GacA without forming heterodimers, either with GacS or with RetS. Enzyme LadS is involved in a genuine phospho relay, which requires both transmitter and receiver LadS domains. LadS signaling ultimately requires the alternative histidine-phosphotransfer domain of GacS. LadS histidine kinase forms, with the GacS/GacA two-component system, a multicomponent signal transduction system with an original phosphorelay cascade, i.e. H1LadS -> D1LadS -> H2GacS -> D2GacA. This highlights an original strategy in which a unique output, i.e. the modulation of sRNA levels, is controlled by a complex multi-sensing network to fine-tune an adapted biofilm and virulence response. In vitro transphosphorylation of GacS H2 domain by LadS histidine kinase, involvement of the H2 domain of the GacS unorthodox histidine kinase in the LadS signaling pathway	Pseudomonas aeruginosa

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Release 2023.1 (January 2023)