24 research outputs found
Mutations in Escherichia coli aceE and ribB genes allow survival of strains defective in the first step of the isoprenoid biosynthesis pathway
A functional 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway is required for isoprenoid biosynthesis and hence survival in Escherichia coli and most other bacteria. In the first two steps of the pathway, MEP is produced from the central metabolic intermediates pyruvate and glyceraldehyde 3-phosphate via 1-deoxy-D-xylulose 5-phosphate (DXP) by the activity of the enzymes DXP synthase (DXS) and DXP reductoisomerase (DXR). Because the MEP pathway is absent from humans, it was proposed as a promising new target to develop new antibiotics. However, the lethal phenotype caused by the deletion of DXS or DXR was found to be suppressed with a relatively high efficiency by unidentified mutations. Here we report that several mutations in the unrelated genes aceE and ribB rescue growth of DXS-defective mutants because the encoded enzymes allowed the production of sufficient DXP in vivo. Together, this work unveils the diversity of mechanisms that can evolve in bacteria to circumvent a blockage of the first step of the MEP pathway
Mutations in escherichia coli aceE and ribB genes allow survival of strains defective in the first step of the isoprenoid biosynthesis pathway
A functional 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway is required for isoprenoid biosynthesis and hence survival in Escherichia coli and most other bacteria. In the first two steps of the pathway, MEP is produced from the central metabolic intermediates pyruvate and glyceraldehyde 3-phosphate via 1-deoxy-D-xylulose 5-phosphate (DXP) by the activity of the enzymes DXP synthase (DXS) and DXP reductoisomerase (DXR). Because the MEP pathway is absent from humans, it was proposed as a promising new target to develop new antibiotics. However, the lethal phenotype caused by the deletion of DXS or DXR was found to be suppressed with a relatively high efficiency by unidentified mutations. Here we report that several mutations in the unrelated genes aceE and ribB rescue growth of DXS-defective mutants because the encoded enzymes allowed the production of sufficient DXP in vivo. Together, this work unveils the diversity of mechanisms that can evolve in bacteria to circumvent a blockage of the first step of the MEP pathway
CMBPol Mission Concept Study: Probing Inflation with CMB Polarization
We summarize the utility of precise cosmic microwave background (CMB)
polarization measurements as probes of the physics of inflation. We focus on
the prospects for using CMB measurements to differentiate various inflationary
mechanisms. In particular, a detection of primordial B-mode polarization would
demonstrate that inflation occurred at a very high energy scale, and that the
inflaton traversed a super-Planckian distance in field space. We explain how
such a detection or constraint would illuminate aspects of physics at the
Planck scale. Moreover, CMB measurements can constrain the scale-dependence and
non-Gaussianity of the primordial fluctuations and limit the possibility of a
significant isocurvature contribution. Each such limit provides crucial
information on the underlying inflationary dynamics. Finally, we quantify these
considerations by presenting forecasts for the sensitivities of a future
satellite experiment to the inflationary parameters.Comment: 107 pages, 14 figures, 17 tables; Inflation Working Group
contribution to the CMBPol Mission Concept Study; v2: typos fixed and
references adde
The First Data Release of the Sloan Digital Sky Survey
The Sloan Digital Sky Survey has validated and made publicly available its
First Data Release. This consists of 2099 square degrees of five-band (u, g, r,
i, z) imaging data, 186,240 spectra of galaxies, quasars, stars and calibrating
blank sky patches selected over 1360 square degrees of this area, and tables of
measured parameters from these data. The imaging data go to a depth of r ~ 22.6
and are photometrically and astrometrically calibrated to 2% rms and 100
milli-arcsec rms per coordinate, respectively. The spectra cover the range
3800--9200 A, with a resolution of 1800--2100. Further characteristics of the
data are described, as are the data products themselves.Comment: Submitted to The Astronomical Journal. 16 pages. For associated
documentation, see http://www.sdss.org/dr
Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries
Abstract
Background
Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres.
Methods
This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries.
Results
In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia.
Conclusion
This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries
Mutations in Escherichia coli aceE and ribB genes allow survival of strains defective in the first step of the isoprenoid biosynthesis pathway
A functional 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway is required for isoprenoid biosynthesis and hence survival in Escherichia coli and most other bacteria. In the first two steps of the pathway, MEP is produced from the central metabolic intermediates pyruvate and glyceraldehyde 3-phosphate via 1-deoxy-D-xylulose 5-phosphate (DXP) by the activity of the enzymes DXP synthase (DXS) and DXP reductoisomerase (DXR). Because the MEP pathway is absent from humans, it was proposed as a promising new target to develop new antibiotics. However, the lethal phenotype caused by the deletion of DXS or DXR was found to be suppressed with a relatively high efficiency by unidentified mutations. Here we report that several mutations in the unrelated genes aceE and ribB rescue growth of DXS-defective mutants because the encoded enzymes allowed the production of sufficient DXP in vivo. Together, this work unveils the diversity of mechanisms that can evolve in bacteria to circumvent a blockage of the first step of the MEP pathway
Mutations in escherichia coli aceE and ribB genes allow survival of strains defective in the first step of the isoprenoid biosynthesis pathway
A functional 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway is required for isoprenoid biosynthesis and hence survival in Escherichia coli and most other bacteria. In the first two steps of the pathway, MEP is produced from the central metabolic intermediates pyruvate and glyceraldehyde 3-phosphate via 1-deoxy-D-xylulose 5-phosphate (DXP) by the activity of the enzymes DXP synthase (DXS) and DXP reductoisomerase (DXR). Because the MEP pathway is absent from humans, it was proposed as a promising new target to develop new antibiotics. However, the lethal phenotype caused by the deletion of DXS or DXR was found to be suppressed with a relatively high efficiency by unidentified mutations. Here we report that several mutations in the unrelated genes aceE and ribB rescue growth of DXS-defective mutants because the encoded enzymes allowed the production of sufficient DXP in vivo. Together, this work unveils the diversity of mechanisms that can evolve in bacteria to circumvent a blockage of the first step of the MEP pathway
DXP levels in cells expressing wild type and mutant <i>aceE</i> and <i>ribB</i> genes.
<p><i>E.coli</i> cells defective in both DXS and DXR were transformed with pCRII-TOPO constructs with the indicated wild type or mutant genes or an empty vector control (Ø). Positive transformants were grown in triplicate for 5 h with MVA, and DXP levels were measured by LC-MS. Mean and standard deviation (n = 3) values are represented relative to the levels in Ø controls.</p
Ocurrence of identified mutations in EcAB4-2 cells.
<p>Ocurrence of identified mutations in EcAB4-2 cells.</p
Complementation of <i>E.coli</i> strains defective in DXS or DXR.
<p>Cells were transformed with pCRII-TOPO constructs and plated on LB medium containing chloramphenicol (for the disruption of chromosomal <i>dxs</i> or <i>dxr</i> genes), kanamycin (for the MVA operon) and ampicillin (for the introduced plasmid). The medium was supplemented (+) or not (−) with MVA as indicated. Plates were incubated at 37°C for 20 h. (A) Position of transformants harboring plasmids with the indicated wild type or mutant (asterisks) genes or an empty vector control (Ø). (B) EcAB4-2 (<i>dxs::CAT</i>) transformants. (C) EcAB4-10 (<i>dxr::CAT</i>) transformants.</p