Proposed Carrier Lipid-binding Site of Undecaprenyl Pyrophosphate Phosphatase from Escherichia coli

Chang, Hung‐Hao; Chou, Chia Cheng; Hsu, Min; Wang, Andrew H.J.

doi:10.1074/jbc.m114.575076

Cited by 36 publications

(93 citation statements)

References 53 publications

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“…In this study, we were particularly interested in two identified mutants that were highly sensitive to bacitracin and had two independent insertions in the same locus, SMU.244, which encodes a homologue of UppP required for the recycling or de novo synthesis of undecaprenyl phosphate (Up; C 55 -P) (Bickford & Nick, 2013;Chang et al, 2014;El Ghachi et al, 2004). Although the roles of UppP in cell wall biosynthesis and bacitracin resistance are well documented in E. coli, UppP homologues and their physiological features in many Gram-positive bacteria, such as S. mutans, are still lacking.…”

Section: Discussionmentioning

confidence: 99%

“…3a, b). We then assayed the enzymic activity of the UppP Sm protein using a phosphatase activity assay according to the methods of Hsu et al (2013) and Chang et al (2014). The results revealed that purified UppP Sm protein displayed a strong activity of catalysing dephosphorylation of Fpp (Fig.…”

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confidence: 99%

“…Region 1 contained a highly conserved, glutamate-rich E/QXXXE motif (aa 17-21) and one histidine (His30) in the first TMS. This region is suggested to be essential for the catalytic activity and substrate binding (Chang et al, 2014). Region 2 contained a conserved PGXSRSXXT motif (aa 170-178) that formed a structural P-loop (phosphate-binding loop) commonly found in many phosphate-binding enzymes (Bickford & Nick, 2013;Chang et al, 2014).…”

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confidence: 99%

“…This region is suggested to be essential for the catalytic activity and substrate binding (Chang et al, 2014). Region 2 contained a conserved PGXSRSXXT motif (aa 170-178) that formed a structural P-loop (phosphate-binding loop) commonly found in many phosphate-binding enzymes (Bickford & Nick, 2013;Chang et al, 2014). Sequence alignments also indicated that UppP Sm proteins from five genome-sequence-completed S. mutans strains, UA159, GS-5, LJ2, ATCC25175 and NCTC11060, shared 99 % of identity in their primary sequences (Fig.…”

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confidence: 99%

“…The phosphatase activity of UppP Sm was determined by the Malachite Green assay using a phosphate colorimetric kit (BioVision) as described previously (Chang et al, 2014;Hsu et al, 2013). The reaction mixture (200 ml) contained 50 mM HEPES, pH 7.0, 150 mM NaCl, 10 mM MgCl 2 , 0.02 % dodecyl maltoside, 35 mM of the substrate farnesyl pyrophosphate (Fpp) (Sigma-Aldrich) and 2 mg purified UppP Sm protein.…”

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Identification and characterization of SMU.244 encoding a putative undecaprenyl pyrophosphate phosphatase protein required for cell wall biosynthesis and bacitracin resistance in Streptococcus mutans

et al. 2015

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Streptococcus mutans in dental biofilms often faces life-threatening threats such as killing by antimicrobial molecules from competing species or from the host. The ability of S. mutans to cope with such threats is crucial for its survival and persistence in dental biofilms. By screening a transposon mutant library, we identified 11 transposon insertion mutants that were sensitive to bacitracin. Two of these mutants, XTn-01 and XTn-03, had an independent insertion in the same locus, SMU.244, which encoded a homologue of undecaprenyl pyrophosphate phosphatase (UppP). In this study, we describe the genetic and phenotypic characterization of SMU.244 in antibiotic resistance. The results revealed that deletion of SMU.244 results in a mutant (XTD244) that is highly sensitive to bacitracin, but confers more resistance to lactococcin G, a class IIb bacteriocin. Introduction of the intact SMU.244 into XTD244 in trans completely restores its resistance to bacitracin and the susceptibility to lactococcin G. The XTD244 was also defective in forming the WT biofilm, although its growth was not significantly affected. Using recombinant protein technology, we demonstrated that the SMU.244-encoded protein displays enzyme activity to catalyse dephosphorylation of the substrate. The lux transcriptional reporter assays showed that S. mutans maintains a moderate level of expression of SMU.244 in the absence of bacitracin, but bacitracin at sub-MICs can further induce its expression. We concluded that SMU.244 encodes an UppP protein that plays important roles in cell wall biosynthesis and bacitracin resistance in S. mutans. The results described here may further our understanding of the molecular mechanisms by which S. mutans copes with antibiotics such as bacitracin.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Identification and characterization of SMU.244 encoding a putative undecaprenyl pyrophosphate phosphatase protein required for cell wall biosynthesis and bacitracin resistance in Streptococcus mutans

et al. 2015

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Chromosomal mutations in Escherichia coli that improve tolerance to nonvolatile side‐products from dilute acid treatment of sugarcane bagasse

Shi

Yomano

York

et al. 2019

Biotech & Bioengineering

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Lignocellulosic biomass provides attractive nonfood carbohydrates for the production of ethanol, and dilute acid pretreatment is a biomass‐independent process for access to these carbohydrates. However, this pretreatment also releases volatile and nonvolatile inhibitors of fermenting microorganisms. To identify unique gene products contributing to sensitivity/tolerance to nonvolatile inhibitors, ethanologenic Escherichia coli strain LY180 was adapted for growth in vacuum‐treated sugarcane bagasse acid hydrolysate (VBHz) lacking furfural and other volatile inhibitors. A mutant, strain AQ15, obtained after approximately 500 generations of growth in VBHz, grew and fermented the sugars in a medium with 50% VBHz. Comparative genome sequence analysis of strains AQ15 and LY180 revealed 95 mutations in strain AQ15. Six of these mutations were also found in strain SL112, an independent inhibitor‐tolerant derivative of strain LY180. Among these six mutations, null mutations in mdh and bacA were identified as contributing factors to VBHz tolerance in strain AQ15, based on the genetic and physiological analysis. The deletion of either gene in strain LY180 increased tolerance to VBHz from approximately 30–50% (vol/vol). Considering the location and physiological role of the two enzymes in the cell, it is likely that the two enzymes contribute to the VBHz sensitivity of ethanologenic E. coli by different mechanisms.

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Bacterial Cell Growth Inhibitors Targeting Undecaprenyl Diphosphate Synthase and Undecaprenyl Diphosphate Phosphatase

et al. 2016

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We synthesized a series of benzoic acids and phenylphosphonic acids and investigated their effects on the growth of Staphylococcus aureus and Bacillus subtilis. One of the most active compounds (7, 5-fluoro-2-(3-(octyloxy)benzamido)benzoic acid, ED50 ~ 0.15 μg/mL) acted synergistically with seven antibiotics known to target bacterial cell wall biosynthesis (a fractional inhibitory concentration index, FICI~0.35, on average) but had indifferent effects in combinations with six non cell-wall biosynthesis inhibitors (FICI~1.45, on average). The most active compounds were found to inhibit two enzymes involved in isoprenoid/bacterial cell wall biosynthesis: undecaprenyl diphosphate synthase (UPPS) and undecaprenyl diphophate phosphatase (UPPP), but not farnesyl diphosphate synthase, and there were good correlations between bacterial cell growth inhibition, UPPS inhibition and UPPP inhibition.

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Proposed Carrier Lipid-binding Site of Undecaprenyl Pyrophosphate Phosphatase from Escherichia coli

Cited by 36 publications

References 53 publications

Identification and characterization of SMU.244 encoding a putative undecaprenyl pyrophosphate phosphatase protein required for cell wall biosynthesis and bacitracin resistance in Streptococcus mutans

Identification and characterization of SMU.244 encoding a putative undecaprenyl pyrophosphate phosphatase protein required for cell wall biosynthesis and bacitracin resistance in Streptococcus mutans

Chromosomal mutations in Escherichia coli that improve tolerance to nonvolatile side‐products from dilute acid treatment of sugarcane bagasse

Bacterial Cell Growth Inhibitors Targeting Undecaprenyl Diphosphate Synthase and Undecaprenyl Diphosphate Phosphatase

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