Publications by authors named "James C Bouwer"

Article Synopsis
  • Human norovirus (HuNV) is a major cause of gastroenteritis globally, primarily from genogroups I and II, and its lifecycle depends on proteins produced during viral replication, including the functional protease-polymerase (ProPol).
  • The study of ProPol's enzymatic activity revealed that it performs similarly or better than the mature polymerase regarding RNA templates, with unique activity on a poly(A) template and varying responses to antiviral compounds.
  • Advanced cryo-electron microscopy was utilized to determine the structure of the ProPol polymerase domain, revealing similarities to the mature polymerase, thus enhancing the understanding of HuNV replication mechanisms.
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Article Synopsis
  • Ring-shaped DNA sliding clamps are crucial for DNA replication and maintenance, requiring clamp loader complexes (CLCs) to open and position them on DNA.
  • The study presents six detailed structures of E. coli CLC with open and closed clamps before and after DNA binding, highlighting key steps in the loading process.
  • Findings reveal that the ATP-bound CLC first 'grabs' the clamp, then opens it enough for DNA to enter, ultimately allowing the clamp to close around the DNA.
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The fine tuning of biological electrical signaling is mediated by variations in the rates of opening and closing of gates that control ion flux through different ion channels. Human ether-a-go-go related gene (HERG) potassium channels have uniquely rapid inactivation kinetics which are critical to the role they play in regulating cardiac electrical activity. Here, we exploit the K sensitivity of HERG inactivation to determine structures of both a conductive and non-conductive selectivity filter structure of HERG.

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SARS-CoV-2 is the third known coronavirus (CoV) that has crossed the animal-human barrier in the last two decades. However, little structural information exists related to the close genetic species within the SARS-related coronaviruses. Here, we present three novel SARS-related CoV spike protein structures solved by single particle cryo-electron microscopy analysis derived from bat (bat SL-CoV WIV1) and civet (cCoV-SZ3, cCoV-007) hosts.

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The Redβ protein of the bacteriophage λ red recombination system is a model annealase which catalyzes single-strand annealing homologous DNA recombination. Here we present the structure of a helical oligomeric annealing intermediate of Redβ, consisting of N-terminal residues 1-177 bound to two complementary 27mer oligonucleotides, determined via cryogenic electron microscopy (cryo-EM) to a final resolution of 3.3 Å.

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In traditional non-flammable electrolytes a trade-off always exists between non-flammability and battery performance. Previous research focused on reducing free solvents and forming anion-derived solid-electrolyte interphase. However, the contribution of solvated anions in boosting the stability of electrolyte has been overlooked.

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Elongation by RNA polymerase is dynamically modulated by accessory factors. The transcription-repair coupling factor (TRCF) recognizes paused/stalled RNAPs and either rescues transcription or initiates transcription termination. Precisely how TRCFs choose to execute either outcome remains unclear.

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Manganese-based Prussian Blue, Na Mn[Fe(CN) ] (MnPB), is a good candidate for sodium-ion battery cathode materials due to its high capacity. However, it suffers from severe capacity decay during battery cycling due to the destabilizing Jahn-Teller distortions it undergoes as Mn is oxidized to Mn . Herein, the structure is stabilized by a thin epitaxial surface layer of nickel-based Prussian Blue (Na Ni[Fe(CN) ]).

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In bacteria, transcription complexes stalled on DNA represent a major source of roadblocks for the DNA replication machinery that must be removed in order to prevent damaging collisions. Gram-positive bacteria contain a transcription factor HelD that is able to remove and recycle stalled complexes, but it was not known how it performed this function. Here, using single particle cryo-electron microscopy, we have determined the structures of Bacillus subtilis RNA polymerase (RNAP) elongation and HelD complexes, enabling analysis of the conformational changes that occur in RNAP driven by HelD interaction.

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ATP synthase produces the majority of cellular energy in most cells. We have previously reported cryo-EM maps of autoinhibited ATP synthase imaged without addition of nucleotide (Sobti et al. 2016), indicating that the subunit ε engages the α, β and γ subunits to lock the enzyme and prevent functional rotation.

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Serial block-face scanning electron microscopy (SBEM) is quickly becoming an important imaging tool to explore three-dimensional biological structure across spatial scales. At probe-beam-electron energies of 2.0 keV or lower, the axial resolution should improve, because there is less primary electron penetration into the block face.

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Energy filtered transmission electron microscopy techniques are regularly used to build elemental maps of spatially distributed nanoparticles in materials and biological specimens. When working with thick biological sections, electron energy loss spectroscopy techniques involving core-loss electrons often require exposures exceeding several minutes to provide sufficient signal to noise. Image quality with these long exposures is often compromised by specimen drift, which results in blurring and reduced resolution.

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Article Synopsis
  • The study introduces a novel direct detection device (DDD) camera for transmission electron microscopy, tested at electron energies of 120 and 200 keV.
  • This DDD camera operates without a scintillator and achieves high signal transfer of up to 65 lines/mm, marking a leap in imaging technology.
  • An image of virus particles is showcased, demonstrating the DDD's superior performance compared to traditional CCD cameras.
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Article Synopsis
  • A prototype direct detection device (DDD) camera system improves spatial resolution and signal to noise ratio in electron microscopy, outperforming traditional CCD systems.
  • The DDD camera features smaller pixel sizes (5 microm) and eliminates the need for a scintillation screen, enhancing image quality.
  • The paper showcases the first large area mosaic image and tomography dataset from the DDD, including an image processing algorithm to correct specimen drift.
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Alignment of the individual images of a tilt series is a critical step in obtaining high-quality electron microscope reconstructions. We report on general methods for producing good alignments, and utilizing the alignment data in subsequent reconstruction steps. Our alignment techniques utilize bundle adjustment.

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Article Synopsis
  • - A new recording device for transmission electron microscopy (TEM) is needed, as current options (film and CCD cameras) are insufficient for high-resolution 3-D particle reconstruction.
  • - An active pixel sensor (APS) array was tested using electron microscopes, revealing that unexpected electron events were caused by backscattering in the silicon substrate during measurements at low beam intensity.
  • - To improve the APS's usability and eliminate unwanted events, it's suggested to thin the silicon substrate to less than 30 microm, which would increase the signal-to-noise ratio to about 10:1 and achieve a spatial resolution of approximately 10 microm in the 200-400 keV energy range.
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We describe the technique and application of energy filtering, automated most-probable loss (MPL) tomography to intermediate voltage electron microscopy (IVEM). We show that for thick, selectively stained biological specimens, this method produces a dramatic increase in resolution of the projections and the computed volumes versus standard unfiltered transmission electron microscopy (TEM) methods. This improvement in resolution is attributed to the reduction of chromatic aberration, which results from the large percentage of inelastic electron-scattering events for thick specimens.

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