Expression, purification, crystallization and preliminary X-ray analysis of the receiver domain of<i>Staphylococcus aureus</i>LytR protein

Shala, Agnesa; Patel, Kevin; Golemi-Kotra, Dasantila; Audette, Gerald F.

doi:10.1107/s1744309113030972

Cited by 2 publications

(3 citation statements)

References 29 publications

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“…Aligned to the loss of PMF within the bacteria, down‐regulation of the LytR protein, part of the LytSR two‐component system, was also observed (Table SII). This system is known to be a membrane‐bound transcription factor that monitors the PMF across the bacterial membrane and its down‐regulation can lead to cidABC and lrgAB operons‐mediated autolytic response within the cell (Shala et al, ).…”

Section: Resultsmentioning

confidence: 99%

Quantitative proteome profiles help reveal efficient xylose utilization mechanisms in solventogenic Clostridium sp. strain BOH3

Basu

Xin

Lim

et al. 2017

Biotech & Bioengineering

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Development of sustainable biobutanol production platforms from lignocellulosic materials is impeded by inefficient five carbon sugar uptake by solventogenic bacteria. The recently isolated Clostridium sp. strain BOH3 is particularly advantaged in this regard as it serves as a model organism which can simultaneously utilize both glucose and xylose for high butanol (>15 g/L) production. Strain BOH3 was, therefore, investigated for its metabolic mechanisms for efficient five carbon sugar uptake using a quantitative proteomics based approach. The proteomics data show that proteins within the CAC1341-1349 operon play a pivotal role for efficient xylose uptake within the cells to produce butanol. Furthermore, up-regulation of key enzymes within the riboflavin synthesis pathway explained that xylose could induce higher riboflavin production capability of the bacteria (e.g., ∼80 mg/L from glucose vs. ∼120 mg/L from xylose). Overall results from the present experimental approach indicated that xylose-fed BOH3 cultures are subjected to high levels of redox stress which coupled with the solvent stress-trigger a sporulation response within the cells earlier than the glucose-fed cultures. The study lays the platform for metabolic engineering strategies in designing organisms for efficient butanol and other value-added chemicals such as riboflavin production. Biotechnol. Bioeng. 2017;114: 1959-1969. © 2017 Wiley Periodicals, Inc.

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Section: Resultsmentioning

confidence: 99%

Quantitative proteome profiles help reveal efficient xylose utilization mechanisms in solventogenic Clostridium sp. strain BOH3

Basu

Xin

Lim

et al. 2017

Biotech & Bioengineering

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“…To test this directly, we examined the ability of LytR to become phosphorylated in the presence of LytS in vitro. Similar to a previous report (Shala et al ., ), we were unable to purify sufficient quantities of full‐length LytS and LytR for these assays, so we purified and tested only the C‐terminal transmitter domain of LytS (designated LytS‐C) and the N‐terminal receiver domain of LytR (designated LytR‐N). In this experiment, LytS‐C was autophosphorylated (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…Overexpression of the LytR receiver domain (amino acids 1-134), designated LytR-N, was performed as outlined by Shala et al (2013). Briefly, overnight cultures of E. coli Rosetta II cells containing the LytR-N and LytR-N D53A overexpression plasmids (pML75 and pML76) were diluted to an OD600 of 0.1 in 100 ml LB (kanamycin 50 mg ml −1 ) in a 500 ml flask.…”

Section: Protein Purification Of Truncated Lyts and Lytrmentioning

confidence: 99%

Identification of the amino acids essential for LytSR‐mediated signal transduction in Staphylococcus aureus and their roles in biofilm‐specific gene expression

Lehman

Bose

Sharma-Kuinkel

et al. 2015

Molecular Microbiology

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Summary Recent studies have demonstrated that expression of the Staphylococcus aureus lrgAB operon is specifically expressed within tower structures during biofilm development. To gain a better understanding of the mechanisms underlying this spatial control of lrgAB expression, we carried out a detailed analysis of the LytSR two-component system. Specifically, a conserved aspartic acid (Asp53) of the LytR response regulator was shown to be the target of phosphorylation, which resulted in enhanced binding to the lrgAB promoter and activation of transcription. In addition, we identified His390 of the LytS histidine kinase as the site of autophosphorylation and Asn394 as a critical amino acid involved in phosphatase activity. Interestingly, LytS-independent activation of LytR was observed during planktonic growth, with acetyl phosphate acting as a phosphodonor to LytR. In contrast, mutations disrupting the function of LytS prevented tower-specific lrgAB expression, providing insight into the physiologic environment within these structures. In addition, over activation of LytR led to increased lrgAB promoter activity during planktonic and biofilm growth and a change in biofilm morphology. Overall, the results of this study are the first to define the LytSR signal transduction pathway, as well as determine the metabolic context within biofilm tower structures that triggers these signaling events.

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Expression, purification, crystallization and preliminary X-ray analysis of the receiver domain ofStaphylococcus aureusLytR protein

Cited by 2 publications

References 29 publications

Quantitative proteome profiles help reveal efficient xylose utilization mechanisms in solventogenic Clostridium sp. strain BOH3

Quantitative proteome profiles help reveal efficient xylose utilization mechanisms in solventogenic Clostridium sp. strain BOH3

Identification of the amino acids essential for LytSR‐mediated signal transduction in Staphylococcus aureus and their roles in biofilm‐specific gene expression

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