Functional mapping of the surface of escherichia coli ribose‐binding protein: Mutations that affect chemotaxis and transport

Binnie, R. A.; Zhang, Huide; Mowbray, Sherry L.; Hermodson, Mark A.

doi:10.1002/pro.5560011212

Cited by 46 publications

(52 citation statements)

References 40 publications

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“…Figure 7 shows the homology model built on the crystal structure templates of ribose binding protein (RBP) and glucose-galactose binding protein (GBP). T211 is predicted to reside on the surface of ChvE, close to the "lips" of two domains in RBP and GBP where mutations that alter the proteinprotein interactions with chemoreceptors and membrane transport proteins have been found (5,7,17). The binding affinity of sugar substrates can also be altered by mutations in this region, even though it is distinct from the buried sugar binding site (34,44).…”

Section: Discussionmentioning

confidence: 99%

Environmental pH Sensing: Resolving the VirA/VirG Two-Component System Inputs for Agrobacterium Pathogenesis

Gao

Lynn

2005

J Bacteriol

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Agrobacterium tumefaciens stands as one of biotechnology's greatest successes, with all plant genetic engineering building on the strategies of this pathogen. By integrating responses to external pHs, phenols, and monosaccharides, this organism mobilizes oncogenic elements to efficiently transform most dicotyledonous plants. We now show that the complex signaling network used to regulate lateral gene transfer can be resolved as individual signaling modules. While pH and sugar perception are coupled through a common pathway, requiring both low pH and sugar for maximal virulence gene expression, various VirA and ChvE alleles can decouple pH and monosaccharide perception. This VirA and ChvE system may represent a common mechanism that underpins external pH perception in prokaryotes, and the use of these simple genetic elements may now be extended to research on specific responses to changes in environmental pH.

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Section: Discussionmentioning

confidence: 99%

Environmental pH Sensing: Resolving the VirA/VirG Two-Component System Inputs for Agrobacterium Pathogenesis

Gao

Lynn

2005

J Bacteriol

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“…A wealth of evidence has demonstrated that some specialized regions located on the surfaces of PSBPs are important for transport and chemotactic functions. In the case of RBP, four distinct regions spanning the N-terminal and C-terminal domains are involved in interaction with its permease (a transport partner), its chemotransducer (a chemotactic partner), or both (5,15). In GBP, one residue was identified as being specifically involved in chemotaxis but not transport (36,49).…”

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confidence: 99%

Molecular Basis of ChvE Function in Sugar Binding, Sugar Utilization, and Virulence in Agrobacterium tumefaciens

Nair

Soto

et al. 2009

J Bacteriol

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ChvE is a chromosomally encoded protein inThe expression of vir genes in A. tumefaciens is activated by plant-released signals, namely, phenolic derivatives, acidic pH, and monosaccharides (for a review, see reference 6), via the combined activities of the periplasmic protein ChvE and the VirA/VirG two-component regulatory system. Upon perception of these plant signals, autophosphorylated VirA, a transmembrane histidine kinase, transfers a phosphoryl group to VirG, a response regulator, and then the phosphorylated VirG activates the expression of vir genes by binding vir boxes in their promoters (8,19,24,31,52).Perception and transduction of the sugar signals is crucial to the virulence of A. tumefaciens: strains lacking ChvE, a chromosomally encoded putative sugar-binding protein, are significantly less virulent than wild-type strains (17,18). Previous studies have shown that, in fact, sugar signaling is neither sufficient for nor absolutely required for vir gene expression. Rather, sugars vastly increase both the sensitivity of VirA to phenol derivatives, such as acetosyringone (AS), and the maximal levels of vir gene expression observed at saturating levels of such compounds (for a review, see reference 26). The periplasmic domain of VirA is required for transduction of the sugar and pH signals (7,8,16,41), whereas the so-called "linker" domain, located in the cytoplasm between the second transmembrane domain and the kinase domain, is required for perception and transduction of the phenolic signals (8,46,47).A working model for the ChvE/sugar/VirA signaling pathway suggests that monosaccharide-bound ChvE interacts with the periplasmic domain of VirA to relieve periplasmic repression, resulting in maximal sensitivity of VirA to phenolic signals (7,11,32,41). However, limited evidence has been presented to reveal how ChvE recognizes monosaccharides and how it interacts with the periplasmic domain of VirA. Shimoda et al. (41) identified a mutant chvE allele [chvE(T211M)] that is able to suppress a sugar-insensitive virA allele [virA(E210V)], thereby restoring the sugar-sensing ability. The suppressing effect of chvE(T211M) was then proposed to be the result of the specific restoration of the capacity of VirA E210V to bind ChvE T211M . However, ChvE T211M also activated wild-type VirA in the absence of sugars (32), suggesting that this mutant may not be a site-specific suppressor of VirA E210V . Based on a homology model of ChvE, a recent study (16) does predict, though, that the residue T211 is located on the surface of the

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“…Plasmid PAC had the wild-type sequences and, for these assays, the genes are expressed constitutively from a low-level pBR322 promoter (Binnie et al, 1992). Thus, pAC/MH-I I is designated wild type for these assays, and the activities of the fusions with the Gly,(G,), thrombin-cleavable (Th), and factor X,-cleavable (FX) bridges are compared to those values in each case.…”

Section: Production and Purification Of Rbsa-c Chimerasmentioning

confidence: 99%

“…The PAC phagemid was constructed in our laboratory by a combination of site-directed mutagenesis (to add or delete restriction endonuclease sites) and ligation of the appropriate fragments to replace the rbsB gene from p4BI (Binnie et al, 1992) with the coding sequences of rbsAC. An Nde I site was incorporated at the 5'-end of rbsA for cloning convenience as was a Hind 111 site immediately downstream of rbsC.…”

Section: Pac and Chimeric Pa-c Derivativesmentioning

confidence: 99%

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The proteins encoded by the rbs operon of escherichia coli: II. Use of chimeric protein constructs to isolate and characterize RbsC

et al. 1996

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Chimeric genes encoding full-length copies of rbsA and rbsC connected by segments coding for short bridge peptides were constructed and expressed in Escherichia coli. Surprisingly, the chimeric genes complemented the strain in which rbsA and rbsC were deleted. The chimeric proteins were overproduced, and the products were purified by affinity chromatography. In order to obtain highly purified protein, a poly-His leader peptide was incorporated so that Ni-chelate affinity chromatography could be employed. The leader peptide and the bridge peptide were designed with factor X,-cleavable sites to permit recovery of the individual RbsA and RbsC protein. A rbsC gene encoding a poly-His leader was also constructed and expressed. Both the chimeric RbsA-C species and the poly-HisRbsC were produced at levels that permitted isolation of the equivalent of milligram quantities of RbsC per liter of culture. This is a substantial increase in amounts from any previous RbsC production vectors. All proteins from the rbs operon have now been overproduced and substantially purified.Keywords: ABC transporters; chimeric proteins; integral membrane proteins; membrane transport proteins; ribose transport operon "ABC transporters" (Higgins, 1992) are defined by their ATPbinding components, which are proteins or domains of a larger protein of roughly 250 amino acid residues in length that show clear amino acid sequence homology between proteins found in species ranging from bacteria to humans. The "ATP-binding cassettes (ABC)" are invariably associated with extremely hydrophobic proteins or domains of the same protein that are also required for a functional transport complex, presumably as membrane-embedded components forming the channel through which the transported ligand passes. In closely related transporters, amino acid sequence homology between the hydrophobic

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Functional mapping of the surface of escherichia coli ribose‐binding protein: Mutations that affect chemotaxis and transport

Cited by 46 publications

References 40 publications

Environmental pH Sensing: Resolving the VirA/VirG Two-Component System Inputs for Agrobacterium Pathogenesis

Environmental pH Sensing: Resolving the VirA/VirG Two-Component System Inputs for Agrobacterium Pathogenesis

Molecular Basis of ChvE Function in Sugar Binding, Sugar Utilization, and Virulence in Agrobacterium tumefaciens

The proteins encoded by the rbs operon of escherichia coli: II. Use of chimeric protein constructs to isolate and characterize RbsC

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