A5065715606- Crystallization and Solubility Studies
- X-ray Diffraction in Crystallography
- RNA and protein synthesis mechanisms
- Crystallography and molecular interactions
- Protein Structure and Dynamics
- DNA and Nucleic Acid Chemistry
- RNA Research and Splicing
- Advanced NMR Techniques and Applications
- Viral Infections and Immunology Research
- RNA modifications and cancer
- Molecular spectroscopy and chirality
- Enzyme Structure and Function
- Electron Spin Resonance Studies
- HIV Research and Treatment
- Diet and metabolism studies
- Cancer, Hypoxia, and Metabolism
- NMR spectroscopy and applications
- Nuclear Physics and Applications
- Bacteriophages and microbial interactions
- X-ray Spectroscopy and Fluorescence Analysis
- Mass Spectrometry Techniques and Applications
- RNA regulation and disease
- Receptor Mechanisms and Signaling
- Metabolism and Genetic Disorders
- RNA Interference and Gene Delivery
Rapt Therapeutics (United States)
2024
Medical University of South Carolina
2006-2017
University of North Carolina at Chapel Hill
2015-2017
David H. Murdock Research Institute
2015-2016
University of Charleston
2014
Scripps Research Institute
2000-2008
Bruker (United States)
2000-2006
University of Toronto
2006
Masaryk University
2006
Max Planck Institute for Biophysical Chemistry
2000-2005
Cancer-associated thrombocytosis has long been linked to poor clinical outcome, but the underlying mechanism is enigmatic. We hypothesized that platelets promote malignancy and resistance therapy by dampening host immunity. show genetic targeting of enhances adoptive T cell cancer. An unbiased biochemical structural biology approach established transforming growth factor β (TGFβ) lactate as major platelet-derived soluble factors obliterate CD4+ CD8+ functions. Moreover, we found are dominant...
In vivo genome correction holds promise for generating durable disease cures; yet, effective stem cell editing remains challenging. this work, we demonstrate that optimized lung-targeting lipid nanoparticles (LNPs) enable high levels of in cells, yielding responses. Intravenously administered gene-editing LNPs activatable tdTomato mice achieved >70% lung editing, sustaining expression >80% epithelial cells 660 days. Addressing cystic fibrosis (CF), NG-ABE8e messenger RNA (mRNA)-sgR553X...
Angles between two interatomic vectors are measured for structure elucidation in solution nuclear magnetic resonance (NMR). The angles can be determined directly by using the effects of dipole-dipole cross-correlated relaxation double-quantum and zero-quantum coherences. rates related to angular geometry without need calibration a Karplus-type curve, as is case scalar coupling measurements, depend only on rotational correlation time molecule an empirical parameter. This makes determination...
A wide range of RNA viruses use programmed -1 ribosomal frameshifting for the production viral fusion proteins. Inspection overlap regions between ORF1a and ORF1b SARS-CoV genome revealed that, similar to all coronaviruses, a frameshift could be used by virus produce protein. Computational analyses signal predicted presence an mRNA pseudoknot containing three double-stranded stem structures rather than two. Phylogenetic showed conservation potential three-stemmed pseudoknots in signals other...
The three-dimensional structure of the complex between calmodulin (CaM) and a peptide corresponding to N-terminal portion CaM-binding domain plasma membrane calcium pump, C20W, has been solved by heteronuclear nuclear magnetic resonance (NMR) spectroscopy. calculation is based on total 1808 intramolecular NOEs 49 intermolecular C20W from heteronuclear-filtered NOESY spectra half-filtered experiment, respectively. Chemical shift differences free Ca(2+)-saturated CaM its with as well reveal...
Hydrogen bond networks stabilize RNA secondary and tertiary structure are thus essentially important for protein recognition.During refinements using either NMR or X-ray techniques, hydrogen bonds were usually inferred indirectly from the proximity of donor acceptor functional groups.Recently, quantitative heteronuclear J(N,N)-HNN COSY experiments introduced that allowed direct identification nitrogen atoms involved in bonds.However, protons base pairing interactions nucleic acids often not...
The molecular determinants of stimulation -1 programmed ribosomal frameshifting (-1 PRF) by RNA pseudoknots are poorly understood. Sugarcane yellow leaf virus (ScYLV) encodes a 28-nt mRNA pseudoknot that promotes PRF between the P1 (protease) and P2 (polymerase) genes in plant luteoviruses. solution structure ScYLV reveals well ordered loop 2 (L2) exhibits continuous stacking A20 through C27 minor groove upper stem 1 (S1), with C25 flipped out triple-stranded stack. Five consecutive triple...
Abstract Messenger RNA encoded signals that are involved in programmed -1 ribosomal frameshifting (-1 PRF) typically two-stemmed hairpin (H)-type pseudoknots (pks). We previously described an unusual three-stemmed pseudoknot from the severe acute respiratory syndrome (SARS) coronavirus (CoV) stimulated PRF. The conserved existence of a third stem–loop suggested important hitherto unknown function. Here we present new information describing structure and function stem SARS pseudoknot....
Enzymatic synthesis methods for the fluorinated 5'-triphosphate analogues 5F-UTP and 5F-CTP have been developed to facilitate 19F-labeling of RNAs biophysical studies. HIV-2 TAR were synthesized using these by in vitro transcription reactions T7 RNA polymerase. The uniform incorporation 5F-U or 5F-C into transcripts does not significantly alter structure thermodynamic stability. Fluorine observed homonuclear 19F−19F heteronuclear 19F−1H NOE experiments providing selective distance...
Long-range scalar 5J(H1',F) couplings were observed in 5-fluoropyrimidine-substituted RNA. We developed a novel S3E-19F-alpha,beta-edited NOESY experiment for quantitation of these long-range couplings, where the J-couplings can be extracted from inspection intraresidual (H1',H6) NOE cross-peaks. Quantum chemical calculations exploited to investigate relation between and conformations around glycosidic bond oligonucleotides. The theoretical dependence on torsion angle chi described by...
Abstract Folate coenzymes are involved in biochemical reactions of one-carbon transfer, and deficiency this vitamin impairs cellular proliferation, migration, survival many cell types. Here, the effect folate restriction on mammary cancer was evaluated using three distinct breast subtypes differing their aggressiveness metastatic potential: noninvasive basal-like (E-Wnt), invasive but minimally claudin-low (M-Wnt), highly (metM-Wntliver) lines, each derived from same pool MMTV-Wnt-1...
Sperm whale myoglobin in which the heme group has been removed (apomyoglobin) unfolds to an equilibrium intermediate form at pH 4 and can be completely unfolded acid low salt conditions. The titration of a pair partially buried histidine side chains, His24 His119, is particularly important for acid-induced formation from native apomyoglobin. Modifying recently introduced 1H−15N HNN-COSY nuclear magnetic resonance (NMR) experiment (Dingley, A. J.; Grzesiek, S. J. Am. Chem. Soc. 1998, 120,...
The production of isotopically labeled RNA remains critical to current NMR structural studies. One approach obtain simple spectra is label with a nucleus that not naturally occurring in RNA. Fluorine-19 can serve as sensitive site-specific probe upon incorporation into Here we report the efficient vitro enzymatic synthesis 2-fluoroadenosine-5'-triphosphate and its HIV-2 transactivation region (TAR) by DNA template-directed transcription using phage T7 polymerase. We provide unequivocal...
The translation of the unspliced and partially spliced viral mRNAs that encode late, structural proteins HIV-1 depends on viral-protein Rev. Oligomeric binding Rev to response element (RRE) in these promotes their export from nucleus thus controls expression. Here, we compared effects hydrophobic hydrophilic mutations within oligomerization domain using assays for oligomeric RNA binding, protein structure, nucleus. alone does not correlate well with transport activity subset mutants....
While it is recognized that protein side chains undergo large reorientations, very little known about the extent and nature of motions within backbone. These can be studied by chemical shift anisotropy (CSA)−dipolar cross-correlated relaxation rates, interpretation which relies on prior knowledge CSA principal axis values directions. Alternatively, dipolar−dipolar rates used. Here we propose a CT-HNCO experiment to measure sum two C'Cα/NCα dipolar−NHN/C'HN dipolar rates. At same time gives...