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Evaluation Of Fusobacterium Nucleatum Enoyl-Acp Reductase (Fabk) As A Narrow-Spectrum Drug Target, Jacob T Rutherford, Kristiana Avad, Chetna Dureja, Krissada Norseeda, Bibek Gc, Chenggang Wu, Dianqing Sun, Kirk E Hevener, Julian G Hurdle
Evaluation Of Fusobacterium Nucleatum Enoyl-Acp Reductase (Fabk) As A Narrow-Spectrum Drug Target, Jacob T Rutherford, Kristiana Avad, Chetna Dureja, Krissada Norseeda, Bibek Gc, Chenggang Wu, Dianqing Sun, Kirk E Hevener, Julian G Hurdle
Journal Articles
Fusobacterium nucleatum, a pathobiont inhabiting the oral cavity, contributes to opportunistic diseases, such as periodontal diseases and gastrointestinal cancers, which involve microbiota imbalance. Broad-spectrum antimicrobial agents, while effective against F. nucleatum infections, can exacerbate dysbiosis. This necessitates the discovery of more targeted narrow-spectrum antimicrobial agents. We therefore investigated the potential for the fusobacterial enoyl-ACP reductase II (ENR II) isoenzyme FnFabK (C4N14_ 04250) as a narrow-spectrum drug target. ENRs catalyze the rate-limiting step in the bacterial fatty acid synthesis pathway. Bioinformatics revealed that of the four distinct bacterial ENR isoforms, F. nucleatum specifically encodes FnFabK. Genetic studies revealed …
Optimization Of The Antifungal Properties Of The Bacterial Peptide Entv By Variant Analysis, Shantanu Guha, Shane A Cristy, Giuseppe Buda De Cesare, Melissa R Cruz, Michael C Lorenz, Danielle A Garsin
Optimization Of The Antifungal Properties Of The Bacterial Peptide Entv By Variant Analysis, Shantanu Guha, Shane A Cristy, Giuseppe Buda De Cesare, Melissa R Cruz, Michael C Lorenz, Danielle A Garsin
Journal Articles
Fungal resistance to commonly used medicines is a growing public health threat, and there is a dire need to develop new classes of antifungals. We previously described a peptide produced by Enterococcus faecalis, EntV, that restricts Candida albicans to a benign form rather than having direct fungicidal activity. Moreover, we showed that one 12-amino acid (aa) alpha helix of this peptide retained full activity, with partial activity down to the 10aa alpha helix. Using these peptides as a starting point, the current investigation sought to identify the critical features necessary for antifungal activity and to screen for new variants …
Quorum-Sensing, Magdalena Podkowik, Andrew I Perault, Gregory Putzel, Andrew Pountain, Jisun Kim, Ashley L Dumont, Erin E Zwack, Robert J Ulrich, Theodora K Karagounis, Chunyi Zhou, Andreas F Haag, Julia Shenderovich, Gregory A Wasserman, Junbeom Kwon, John Chen, Anthony R Richardson, Jeffrey N Weiser, Carla R Nowosad, Desmond S Lun, Dane Parker, Alejandro Pironti, Xilin Zhao, Karl Drlica, Itai Yanai, Victor J Torres, Bo Shopsin
Quorum-Sensing, Magdalena Podkowik, Andrew I Perault, Gregory Putzel, Andrew Pountain, Jisun Kim, Ashley L Dumont, Erin E Zwack, Robert J Ulrich, Theodora K Karagounis, Chunyi Zhou, Andreas F Haag, Julia Shenderovich, Gregory A Wasserman, Junbeom Kwon, John Chen, Anthony R Richardson, Jeffrey N Weiser, Carla R Nowosad, Desmond S Lun, Dane Parker, Alejandro Pironti, Xilin Zhao, Karl Drlica, Itai Yanai, Victor J Torres, Bo Shopsin
Journal Articles
The agr quorum-sensing system links Staphylococcus aureus metabolism to virulence, in part by increasing bacterial survival during exposure to lethal concentrations of H2O2, a crucial host defense against S. aureus. We now report that protection by agr surprisingly extends beyond post-exponential growth to the exit from stationary phase when the agr system is no longer turned on. Thus, agr can be considered a constitutive protective factor. Deletion of agr resulted in decreased ATP levels and growth, despite increased rates of respiration or fermentation at appropriate oxygen tensions, suggesting that Δagr cells undergo a shift towards a hyperactive metabolic …
Chimeric Systems Composed Of Swapped Tra Subunits Between Distantly-Related F Plasmids Reveal Striking Plasticity Among Type Iv Secretion Machines, Kouhei Kishida, Yang Grace Li, Natsumi Ogawa-Kishida, Pratick Khara, Abu Amar M Al Mamun, Rachel E Bosserman, Peter J Christie
Chimeric Systems Composed Of Swapped Tra Subunits Between Distantly-Related F Plasmids Reveal Striking Plasticity Among Type Iv Secretion Machines, Kouhei Kishida, Yang Grace Li, Natsumi Ogawa-Kishida, Pratick Khara, Abu Amar M Al Mamun, Rachel E Bosserman, Peter J Christie
Journal Articles
Bacterial type IV secretion systems (T4SSs) are a versatile family of macromolecular translocators, collectively able to recruit diverse DNA and protein substrates and deliver them to a wide range of cell types. Presently, there is little understanding of how T4SSs recognize substrate repertoires and form productive contacts with specific target cells. Although T4SSs are composed of a number of conserved subunits and adopt certain conserved structural features, they also display considerable compositional and structural diversity. Here, we explored the structural bases underlying the functional versatility of T4SSs through systematic deletion and subunit swapping between two conjugation systems encoded by the …
Important Features For Protein Foldings In Two Acyl Carrier Proteins From Enterococcus Faecalis, Seoyeong Yoo, Jiwon Yeon, Eunhee Kim, Yangmee Kim
Important Features For Protein Foldings In Two Acyl Carrier Proteins From Enterococcus Faecalis, Seoyeong Yoo, Jiwon Yeon, Eunhee Kim, Yangmee Kim
Journal Articles
The emergence of multi-drug resistant Enterococcus faecalis raises a serious threat to global public health. E. faecalis is a gram-positive intestinal commensal bacterium found in humans. E. faecalis can endure extreme environments such as high temperature, pressure, and high salt, which facilitates them to cause infection in hospitals. E. faecalis has two acyl carrier proteins, AcpA (EfAcpA) in de novo fatty acid synthesis (FAS) and AcpB (EfAcpB) which utilizes exogenous fatty acids. Previously, we determined the tertiary structures of these two ACPs and investigated their structure-function relationships. Solution structures revealed that overall folding of these two ACPs is similar to …
Construction And Characterization Of Functional Ftsa Sandwich Fusions For Studies Of Ftsa Localization And Dynamics During Escherichia Coli Cell Division, Todd A Cameron, William Margolin
Construction And Characterization Of Functional Ftsa Sandwich Fusions For Studies Of Ftsa Localization And Dynamics During Escherichia Coli Cell Division, Todd A Cameron, William Margolin
Journal Articles
FtsA, a homolog of actin, is essential for cell division of Escherichia coli and is widely conserved among many bacteria. FtsA helps to tether polymers of the bacterial tubulin homolog FtsZ to the cytoplasmic membrane as part of the cytokinetic Z ring. GFP fusions to FtsA have illuminated FtsA’s localization in live E. coli, but these fusions have not been fully functional and required the presence of the native FtsA. Here, we characterize “sandwich” fusions of E. coli FtsA to either mCherry or msfGFP that are functional for cell division and exhibit fluorescent rings at midcell that persist throughout …
Overlapping And Distinct Functions Of The Paralogous Pagr Regulators Of Bacillus Anthracis, Ileana D Corsi, Theresa M Koehler
Overlapping And Distinct Functions Of The Paralogous Pagr Regulators Of Bacillus Anthracis, Ileana D Corsi, Theresa M Koehler
Journal Articles
The Bacillus anthracis pagA gene, encoding the protective antigen component of anthrax toxin, is part of a bicistronic operon on pXO1 that codes for its own repressor, PagR1. In addition to the pagAR1 operon, PagR1 regulates sap and eag, two chromosome genes encoding components of the surface layer, a mounting structure for surface proteins involved in virulence. Genomic studies have revealed a PagR1 paralog, PagR2, encoded by a gene on pXO2. The amino acid sequences of the paralogues are 71% identical and show similarity to the ArsR family of transcription regulators. We determined that the expression of either rPagR1 …
Short Signature Rpob Gene Sequence To Differentiate Species In Mycobacterium Abscessus Group, Jian R Bao, Kileen L Shier, Ronald N Master, Robert S Jones, Richard B Clark
Short Signature Rpob Gene Sequence To Differentiate Species In Mycobacterium Abscessus Group, Jian R Bao, Kileen L Shier, Ronald N Master, Robert S Jones, Richard B Clark
Journal Articles
Mycobacterium abscessus group (MAG) are rapidly growing acid-fast bacteria that consist of three closely related species: M. abscessus (Ma), M. bolletii (Mb), and M. massiliense (Mm). Differentiation of these species can be difficult but is increasingly requested owing to recent infectious outbreaks and their differential drug resistance. We developed a novel and rapid pyrosequencing method using short signature sequences (35 to 45 bp) at a hypervariable site in the rpoB gene to differentiate the three MAG species, along with M. chelonae (Mc), and M. immunogenum (Mi). This method was evaluated …
Incfv Plasmid Ped208: Sequence Analysis And Evidence For Translocation Of Maintenance/Leading Region Proteins Through Diverse Type Iv Secretion Systems., Abu Amar M Al Mamun, Kimberly Kissoon, Kouhei Kishida, William C Shropshire, Blake Hanson, Peter J Christie
Incfv Plasmid Ped208: Sequence Analysis And Evidence For Translocation Of Maintenance/Leading Region Proteins Through Diverse Type Iv Secretion Systems., Abu Amar M Al Mamun, Kimberly Kissoon, Kouhei Kishida, William C Shropshire, Blake Hanson, Peter J Christie
Journal Articles
Two phylogenetically distantly-related IncF plasmids, F and pED208, serve as important models for mechanistic and structural studies of F-like type IV secretion systems (T4SSFs) and F pili. Here, we present the pED208 sequence and compare it to F and pUMNF18, the closest match to pED208 in the NCBI database. As expected, gene content of the three cargo regions varies extensively, although the maintenance/leading regions (MLRs) and transfer (Tra) regions also carry novel genes or motifs with predicted modulatory effects on plasmid stability, dissemination and host range. By use of a Cre recombinase assay for translocation (CRAfT), we recently reported that …
Study Of Polytopic Membrane Protein Topological Organization As A Function Of Membrane Lipid Composition, Mikhail Bogdanov, Philip N Heacock, William Dowhan
Study Of Polytopic Membrane Protein Topological Organization As A Function Of Membrane Lipid Composition, Mikhail Bogdanov, Philip N Heacock, William Dowhan
Journal Articles
A protocol is described using lipid mutants and thiol-specific chemical reagents to study lipid-dependent and host-specific membrane protein topogenesis by the substituted-cysteine accessibility method as applied to transmembrane domains (SCAM). SCAM is adapted to follow changes in membrane protein topology as a function of changes in membrane lipid composition. The strategy described can be adapted to any membrane system.
Clinical And Microbiological Aspects Of Linezolid Resistance Mediated By The Cfr Gene Encoding A 23s Rrna Methyltransferase, Cesar A Arias, Martha Vallejo, Jinnethe Reyes, Diana Panesso, Jaime Moreno, Elizabeth Castañeda, Maria V Villegas, Barbara E Murray, John P Quinn
Clinical And Microbiological Aspects Of Linezolid Resistance Mediated By The Cfr Gene Encoding A 23s Rrna Methyltransferase, Cesar A Arias, Martha Vallejo, Jinnethe Reyes, Diana Panesso, Jaime Moreno, Elizabeth Castañeda, Maria V Villegas, Barbara E Murray, John P Quinn
Journal Articles
The cfr (chloramphenicol-florfenicol resistance) gene encodes a 23S rRNA methyltransferase that confers resistance to linezolid. Detection of linezolid resistance was evaluated in the first cfr-carrying human hospital isolate of linezolid and methicillin-resistant Staphylococcus aureus (designated MRSA CM-05) by dilution and diffusion methods (including Etest). The presence of cfr was investigated in isolates of staphylococci colonizing the patient's household contacts and clinical isolates recovered from patients in the same unit where MRSA CM-05 was isolated. Additionally, 68 chloramphenicol-resistant Colombian MRSA isolates recovered from hospitals between 2001 and 2004 were screened for the presence of the cfr gene. In addition to erm(B), …
Functional Taxonomy Of Bacterial Hyperstructures, Vic Norris, Tanneke Den Blaauwen, Armelle Cabin-Flaman, Roy H Doi, Rasika Harshey, Laurent Janniere, Alfonso Jimenez-Sanchez, Ding Jun Jin, Petra Anne Levin, Eugenia Mileykovskaya, Abraham Minsky, Milton Saier, Kirsten Skarstad
Functional Taxonomy Of Bacterial Hyperstructures, Vic Norris, Tanneke Den Blaauwen, Armelle Cabin-Flaman, Roy H Doi, Rasika Harshey, Laurent Janniere, Alfonso Jimenez-Sanchez, Ding Jun Jin, Petra Anne Levin, Eugenia Mileykovskaya, Abraham Minsky, Milton Saier, Kirsten Skarstad
Journal Articles
The levels of organization that exist in bacteria extend from macromolecules to populations. Evidence that there is also a level of organization intermediate between the macromolecule and the bacterial cell is accumulating. This is the level of hyperstructures. Here, we review a variety of spatially extended structures, complexes, and assemblies that might be termed hyperstructures. These include ribosomal or "nucleolar" hyperstructures; transertion hyperstructures; putative phosphotransferase system and glycolytic hyperstructures; chemosignaling and flagellar hyperstructures; DNA repair hyperstructures; cytoskeletal hyperstructures based on EF-Tu, FtsZ, and MreB; and cell cycle hyperstructures responsible for DNA replication, sequestration of newly replicated origins, segregation, compaction, and …
Vancomycin-Resistant Enterococcus Faecalis Endocarditis: Linezolid Failure And Strain Characterization Of Virulence Factors, Constantine Tsigrelis, Kavindra V Singh, Thais D Coutinho, Barbara E Murray, Larry M Baddour
Vancomycin-Resistant Enterococcus Faecalis Endocarditis: Linezolid Failure And Strain Characterization Of Virulence Factors, Constantine Tsigrelis, Kavindra V Singh, Thais D Coutinho, Barbara E Murray, Larry M Baddour
Journal Articles
Infective endocarditis due to vancomycin-resistant (VR) Enterococcus faecalis has only rarely been reported. We report a case of VR E. faecalis endocarditis that failed to respond to linezolid therapy, outline the virulence traits of the isolate, and review previously published cases of VR E. faecalis endocarditis.