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Articles 61 - 90 of 229
Full-Text Articles in Microbiology
Posttranslational Modification And Protein Disorder Regulate Protein-Protein Interactions And Dna Binding Specificity Of P53, Robin Levy
USF Tampa Graduate Theses and Dissertations
p53 is an intrinsically disordered transcription factor that suppresses tumor development by arresting the cell cycle and promoting DNA repair. p53 deletions or mutations can lead to cancer due to the inability of cells to respond to stress. The protein levels and post-translational modification state of p53 changes in response to cellular stress like DNA damage. Previous studies have shown that p53 can undergo coupled folding and binding with the E3 ubiquitin ligase, Mdm2, and the histone deacetylase, p300. In normal cells, p53 is kept at a low level by Mdm2, which marks it with ubiquitin, targeting p53 for proteasome …
Spatial And Temporal Determinants Associated With Eastern Equine Encephalitis Virus Activity In Florida, Kristi M. Miley
Spatial And Temporal Determinants Associated With Eastern Equine Encephalitis Virus Activity In Florida, Kristi M. Miley
USF Tampa Graduate Theses and Dissertations
Eastern Equine Encephalitis virus (EEEV) is considered the most pathogenic mosquito-borne illness in the United States. Human mortality has been estimated to range between 35 to 75%, with horses experiencing mortality rates greater than 90%. A large number of EEEV cases occur in Florida. Though we have come a long way since the first human infection was identified in the 1930s, there is still much to learn regarding the virus’s ability to maintain transmission year-round in Florida. Phylogenic studies support that Florida may serve as the geographic reservoir for EEEV. This research investigated spatiotemporal and ecological variables associated with risk …
The Impact Of Sample Processing And Media Chemistry On The Culturable Diversity Of Bacteria Isolated From A Cave, Katey E. Bender, Katelyn Glover, Alexander Archey, Hazel A. Barton
The Impact Of Sample Processing And Media Chemistry On The Culturable Diversity Of Bacteria Isolated From A Cave, Katey E. Bender, Katelyn Glover, Alexander Archey, Hazel A. Barton
International Journal of Speleology
Although molecular approaches can identify members of microbial communities in the environment, genomic information does not necessarily correlate with environmental phenotype. Understanding functional roles can be done by cultivating representative species, yet the culturablility of bacteria from caves remains low, at 0.02%, limiting our understanding of microbial community interactions and processes. We have investigated several factors influencing culturability of bacteria from a single sample location in Maxwelton Sink Cave, WV, USA. Extended incubation of inoculated plates showed a significant increase in colony counts from two to four weeks, indicating that extended incubations increase culturability. There were no significant differences in …
Did You Wash Your Caving Suit? Cavers’ Role In The Potential Spread Of Pseudogymnoascus Destructans, The Causative Agent Of White-Nose Disease, Violeta Zhelyazkova, Antonia Hubancheva, Georgi Radoslavov, Nia Toshkova, Sebastien J. Puechmaille
Did You Wash Your Caving Suit? Cavers’ Role In The Potential Spread Of Pseudogymnoascus Destructans, The Causative Agent Of White-Nose Disease, Violeta Zhelyazkova, Antonia Hubancheva, Georgi Radoslavov, Nia Toshkova, Sebastien J. Puechmaille
International Journal of Speleology
White-Nose Disease (WND) has killed millions of hibernating bats in the US and Canada. Its causative agent, the fungus Pseudogymnoascus destructans was introduced to North America, but is native to Europe and Asia, where it is not associated with mass mortality. Although it is nearly impossible to eradicate an emerging wildlife disease, research on P. destructans spread mechanisms can aid in prevention of new introductions and development of better environmental management strategies. It is of particular importance to quantify the potential role of people visiting caves (cavers, tourists, bat researchers, etc.) whom inadvertently move P. destructans spores between sites, and …
Microbial Community Structures In Three Bahamian Blue Holes, Meghan J. Gordon
Microbial Community Structures In Three Bahamian Blue Holes, Meghan J. Gordon
USF Tampa Graduate Theses and Dissertations
This study used 16S rDNA metagenomics and water chemistry to conduct an examination of microbial community dynamics and biogeochemistry in three physically adjacent, sunlit blue holes with variable hydrologic regimes on San Salvador Island, Bahamas. Church and Watling’s Blue Holes are holomictic with relatively clear waters, while Inkwell Blue Hole hosts density stratification and waters stained brown with tannins. Based on water color and clarity and physicochemical profiles, I hypothesized Church and Watling’s Blue Holes would be dominated by oxigenic photoautotrophs, and that the bottom layer of Inkwell would be characterized by euxinic (anoxic and sulfidic) conditions and host primarily …
Surface Runoff Alters Cave Microbial Community Structure And Function, Madison Davis, Maria A. Messina, Giuseppe Nicolosi, Salvatore Petralia, Melvin D. Baker, Christiana K. S. Mayne, Chelsea M. Dinon, Christina J. Moss, Bogdan P. Onac, James R. Garey
Surface Runoff Alters Cave Microbial Community Structure And Function, Madison Davis, Maria A. Messina, Giuseppe Nicolosi, Salvatore Petralia, Melvin D. Baker, Christiana K. S. Mayne, Chelsea M. Dinon, Christina J. Moss, Bogdan P. Onac, James R. Garey
Molecular Biosciences Faculty Publications
Caves formed by sulfuric acid dissolution have been identified worldwide. These caves can host diverse microbial communities that are responsible for speleogenesis and speleothem formation. It is not well understood how microbial communities change in response to surface water entering caves. Illumina 16S rRNA sequencing and bioinformatic tools were used to determine the impact of surface water on the microbial community diversity and function within a spring pool found deep in the Monte Conca Cave system in Sicily, Italy. Sulfur oxidizers comprised more than 90% of the microbial community during the dry season and were replaced by potential anthropogenic contaminants …
To Mid-Cell And Beyond: Characterizing The Roles Of Gpsb And Ypsa In Cell Division Regulation In Gram-Positive Bacteria, Robert S. Brzozowski
To Mid-Cell And Beyond: Characterizing The Roles Of Gpsb And Ypsa In Cell Division Regulation In Gram-Positive Bacteria, Robert S. Brzozowski
USF Tampa Graduate Theses and Dissertations
The bacterial cell division protein FtsZ is a tubulin homolog that forms a ring-like structure at the site of cell division in most bacterial species. There it acts as a scaffold, aiding in the recruitment of other divisome proteins to the site of cell division. Furthermore, studies focusing on the role of FtsZ treadmilling and septal peptidoglycan synthesis implicates that FtsZ plays a direct role in the ultimate closure of the division septum. Thus, many studies in the field of bacterial cell division have focused on FtsZ in terms of its spatial and temporal regulation as well as its ability …
Targeting The Eskape Pathogens By Botanical And Microbial Approaches, Emily Dilandro
Targeting The Eskape Pathogens By Botanical And Microbial Approaches, Emily Dilandro
USF Tampa Graduate Theses and Dissertations
There is a vital need to find new clinical treatment options to combat ESKAPE pathogen infections. Nature has thus far been the most fruitful at providing antimicrobial compounds, which have been derived from a plethora of sources. Ranging from plants to microbial communities, these organisms create chemical compounds that are used as defense mechanisms against invasive or encroaching organisms and confer the producers with competitive advantages. In this study, cinnamaldehyde was investigated as a botanical approach to finding active antimicrobial compounds that inhibit the ESKAPE pathogens. Here, we show that all the ESKAPE pathogens are inhibited by cinnamaldehyde concentrations between …
The Ecology And Conservation Of An Urban Karst Subterranean Estuary, Robert J. Scharping
The Ecology And Conservation Of An Urban Karst Subterranean Estuary, Robert J. Scharping
USF Tampa Graduate Theses and Dissertations
Sulphur Springs is an artesian spring and part of the limestone karst topography of metropolitan Tampa, Florida, USA. Underlying this spring is an extensive cave system that hosts saltwater vents and conspicuous microbial biofilm communities. For decades, water from Sulphur Springs has been extracted and used to supplement public water supply and dry season flows in the Hillsborough River Estuary. This dissertation describes research conducted at Sulphur Springs to determine the impacts of urban land use on the underlying cave and downstream estuary ecosystems, and presents the use of this system as a model to fill certain ecological knowledge gaps. …
Active Growth Of Non-Hydrothermal Subaqueous And Subaerial Barite (Baso4) Speleothems In Lechuguilla Cave (New Mexico, Usa), Max Wisshak, Hazel A. Barton, Katey E. Bender, Harvey R. Duchene
Active Growth Of Non-Hydrothermal Subaqueous And Subaerial Barite (Baso4) Speleothems In Lechuguilla Cave (New Mexico, Usa), Max Wisshak, Hazel A. Barton, Katey E. Bender, Harvey R. Duchene
International Journal of Speleology
Barite (BaSO4) speleothems have been reported from caves around the globe and interpreted to have chiefly formed in phreatic, hypogene, hydrothermal settings. Here we report two contrasting types of barite speleothems (bluish tabular crystals in a shallow pool and actively dripping greenish stalactites), which today form at lower temperatures in the non-hydrothermal and vadose environment of Lechuguilla Cave, New Mexico, USA. Scanning electron microscopy analysis, along with energy- and wavelength-dispersive X-ray spectroscopy (EDS, WDS), as well as X-ray diffraction (XRD), characterize the habit and chemical composition as barite. Fractionation of the minor element calcium is related to growth …
Survey Of Histoplasma Capsulatum In Bat Guano And Status Of Histoplasmosis In Slovenia, Central Europe, Janez Mulec, Saša Simčič, Tadeja Kotar, Romina Kofol, Sanja Stopinšek
Survey Of Histoplasma Capsulatum In Bat Guano And Status Of Histoplasmosis In Slovenia, Central Europe, Janez Mulec, Saša Simčič, Tadeja Kotar, Romina Kofol, Sanja Stopinšek
International Journal of Speleology
There have been increasing reports on the presence of Histoplasma capsulatum in some European countries. The study investigated the presence of Histoplasma in bat guanos, speleologists with records of visiting Histoplasma-endemic regions and patients with histoplasmosis. A commercial ALPHA Histoplasma Antigen enzyme immunoassay was tested as an alternative methodology to detect Histoplasma in environment and compared with polymerase chain reaction (PCR) assays. The presence of Histoplasma antigen in bat guanos was not confirmed by PCR. Among 14 healthy speleologists, two were indicated as having the Histoplasma antigen in urine, but expressed negative PCR-specific results for the Histoplasma antigen. Five …
Draft Genome Sequence Of Verrucosispora Sp. Strain Cwr15, Isolated From A Gulf Of Mexico Sponge, Sarah J. Kennedy, Eleonora Cella, Mohammad Jubair, Taj Azarian, Bill J. Baker, Lindsey N. Shaw
Draft Genome Sequence Of Verrucosispora Sp. Strain Cwr15, Isolated From A Gulf Of Mexico Sponge, Sarah J. Kennedy, Eleonora Cella, Mohammad Jubair, Taj Azarian, Bill J. Baker, Lindsey N. Shaw
Molecular Biosciences Faculty Publications
Here, we present the draft genome sequence of Verrucosispora sp. strain CWR15, a bacterial symbiont of a Gulf of Mexico sponge. The genome consists of 35 contigs encoding 5,840 genes. The genome is the basis for future study and presents an underexplored taxonomy and biosynthetic potential.
Mir146a-Mediated Targeting Of Fancm During Inflammation Compromises Genome Integrity, Devakumar Sundaravinayagam, Hye Rim Kim, Tingting Wu, Hyun Hee Kim, Semo Jun, Jeong-Heon Cha, Younghoon Kee, Ho Jin You, Jung-Hee Lee
Mir146a-Mediated Targeting Of Fancm During Inflammation Compromises Genome Integrity, Devakumar Sundaravinayagam, Hye Rim Kim, Tingting Wu, Hyun Hee Kim, Semo Jun, Jeong-Heon Cha, Younghoon Kee, Ho Jin You, Jung-Hee Lee
Molecular Biosciences Faculty Publications
Inflammation is a potent inducer of tumorigenesis. Increased DNA damage or loss of genome integrity is thought to be one of the mechanisms linking inflammation and cancer development. It has been suggested that NF-κB-induced microRNA-146 (miR146a) may be a mediator of the inflammatory response. Based on our initial observation that miR146a overexpression strongly increases DNA damage, we investigated its potential role as a modulator of DNA repair. Here, we demonstrate that FANCM, a component in the Fanconi Anemia pathway, is a novel target of miR146a. miR146a suppressed FANCM expression by directly binding to the 3’ untranslated region of the gene. …
Cornus Officinalis Promotes Igfbp2 And Autophagy In Human 1.1b4 Pancreatic Cell Line As Revealed By Employing A Global Proteomic Approach Via Mass Spectrometry, Arielle Elizabeth Tawfik, Jennifer Guergues, Stanley M. Stevens Jr, Brant Roger Burkhardt
Cornus Officinalis Promotes Igfbp2 And Autophagy In Human 1.1b4 Pancreatic Cell Line As Revealed By Employing A Global Proteomic Approach Via Mass Spectrometry, Arielle Elizabeth Tawfik, Jennifer Guergues, Stanley M. Stevens Jr, Brant Roger Burkhardt
Molecular Biosciences Faculty Publications
Type 1 diabetes (T1D) results in the loss of pancreatic beta cells and subsequent loss of insulin production. Exogenous insulin is the only effective treatment, but there is still no cure or interventional therapy available to inhibit progression of T1D. Successful T1D interventional therapy must protect pancreatic beta cells from autoimmunity while enhancing beta cell survival and function. Our data suggests Cornus officinalis (CO) may be a candidate for interventional therapy to protect pancreatic beta cells from autoimmune attack and increase their function. CO has been used in traditional Chinese medicine (TCM) for over 2,000 years and has shown characteristics …
Screening The Cultivable Cave Microbial Mats For The Production Of Antimicrobial Compounds And Antibiotic Resistance, Jerneja Ambrožič Avguštin, Patricia Petrič, Lejla Pašić
Screening The Cultivable Cave Microbial Mats For The Production Of Antimicrobial Compounds And Antibiotic Resistance, Jerneja Ambrožič Avguštin, Patricia Petrič, Lejla Pašić
International Journal of Speleology
The current work extends the phenotypic characterization of a bacterial culture collection obtained from white, yellow, grey or pink microbial cave wall colonies that are common in the caves of Slovenian Karst. We have determined antibiotic resistance to 22 natural and synthetic antibiotics in 69 isolates from the microbial mats. Thirty-eight isolates (52%) were resistant to 1-5 antibiotics; another 27 isolates (37%) were resistant to 6-10 antibiotics; and 7 isolates (0.1%) were resistant to 11-17 antibiotics. We screened for production of antimicrobial compounds by growing cave isolates on five different media and overlaying individual cultures with ten Gram-positive and Gram …
The Role Of Secreted Proteases In Regulating Disease Progression In Staphylococcus Aureus, Brittney D. Gimza
The Role Of Secreted Proteases In Regulating Disease Progression In Staphylococcus Aureus, Brittney D. Gimza
USF Tampa Graduate Theses and Dissertations
Staphylococcus aureus is a highly successful pathogen capable of producing a wealth of virulence factors in the human host. Of note, ten extracellular proteases are produced alongside these virulence factors and play a multifaceted role during infection. They not only cleave host proteins to promote bacterial invasion, immune evasion and survival, but also control disease progression by modulating the stability of self-derived pathogenic determinants. The importance of the secreted proteases modulating virulence factor stability is evidenced by our groups previous finding that a protease-null strain has a substantially increased infectious capacity in a murine model of sepsis; resulting from the …
The Adaptations Of The Sulfur-Oxidizing, Gammaproteobacterium "Candidatus Endoriftia Persephone", Endosymbiont Of The Giant Tubeworm Riftia Pachyptila, To Hydrothermal Vent Habitat Heterogeneity, Juliana M. Leonard
USF Tampa Graduate Theses and Dissertations
The siboglinid tubeworm Riftia pachyptila is a dominant member of the deep-sea megafauna where seawater and hydrothermal vent (HTV) effluent interface and mix. It is one of the fastest growing invertebrates on land or in the sea. It does not have a digestive tract (e.g. mouth, gut, or anus), and is completely dependent on its sulfur-oxidizing endosymbiont, the Gammaproteobacterium “Candidatus Endoriftia persephone” for its nutritional requirements. This association was the first and is the most well studied among chemolithoautotrophic symbioses. “Ca. E. persephone” is a chemolithoautotrophic bacterium that oxidizes sulfide as an electron donor for energy, reduces oxygen as a …
Targeting Bacterial Resistance And Cancer Metastasis: A Structure Based Approach, Kyle Galen Kroeck
Targeting Bacterial Resistance And Cancer Metastasis: A Structure Based Approach, Kyle Galen Kroeck
USF Tampa Graduate Theses and Dissertations
Current research in pharmaceutical development commonly utilizes a profusion of methods in molecular modeling in order to probe intricate biological problems. Many original and promising compounds have been identified and developed by integrating experimental and computational methods. Structural biology utilizes many different research techniques including x-ray crystallography, NMR, and electron microscopy in order to develop molecular models of macromolecules that are of biological interest. Such techniques can be used in conjunction with molecular docking, which utilizes those molecular models in order to target macromolecules of therapeutic interest by computationally analyzing the conformations adopted by ligands upon interaction with a desired …
Design And Implementation Of Degenerate Qpcr/Qrt-Pcr Primers To Detect Microbial Nitrogen Metabolism In Wastewater And Wastewater-Related Samples, Ryan F. Keeley
USF Tampa Graduate Theses and Dissertations
Nitrogen cycling processes can be tracked using quantitative Polymerase Chain Reaction (qPCR) to determine the presence and qReverse Transcriptase-PCR (qRT-PCR) to determine expression of key genes, or ‘biological markers’, for nitrogen metabolism. Nitrification is catalyzed in part, by two enzymes: ammonia monooxygenase (AMO; NH3 NH2OH) and nitrite oxidoreductase (NXR; NO2- NO3-). For denitrification, four enzymes act sequentially: nitrate reductase (NAR/NAP; NO3- NO2-), nitrite reductase (NIR; NO2- NO), nitric oxide reductase (NOR; NO N2O), and nitrous oxide reductase (NOS; N2O N2). A principle of wastewater treatment (WWT) is to remove excess nitrogen by taking advantage of natural nitrogen cycling …
Are Microclimate Conditions In El Malpais National Monument Caves In New Mexico, Usa Suitable For Pseudogymnoascus Growth?, Terry J. Torres-Cruz, Andrea Porras-Alfaro, Nicole A. Caimi, Ogochukwu Nwabologu, Edward W. Strach, Kaitlyn J.H. Read, Jesse M. Young, Debbie C. Buecher, Diana E. Northup
Are Microclimate Conditions In El Malpais National Monument Caves In New Mexico, Usa Suitable For Pseudogymnoascus Growth?, Terry J. Torres-Cruz, Andrea Porras-Alfaro, Nicole A. Caimi, Ogochukwu Nwabologu, Edward W. Strach, Kaitlyn J.H. Read, Jesse M. Young, Debbie C. Buecher, Diana E. Northup
International Journal of Speleology
White-nose syndrome (WNS) is a bat disease caused by the fungal pathogen Pseudogymnoascus destructans, which thrives in cold and very humid environments where bats frequently hibernate. Conidia of Pseudogymnoascus species are often documented on bats prior to the onset of WNS, but characterization of high-risk areas defined by microclimate cave conditions have been lacking. Investigating the occurrence of this fungal genus and appropriate environmental conditions to support P. destructans in southwestern U.S. caves is key to understanding the sites most likely to be impacted by WNS. Microclimate conditions in ten caves at El Malpais (ELMA) National Monument in New …
A Small Rna And Dna Binding Protein Contribute To Biofilm Development In Bartonella Henselae, Udoka Okaro
A Small Rna And Dna Binding Protein Contribute To Biofilm Development In Bartonella Henselae, Udoka Okaro
USF Tampa Graduate Theses and Dissertations
A biofilm, which is associated with 80% of chronic infections in humans, is formed when bacteria aggregate, attach to a substrate and secrete a matrix protecting the bacteria from host cell defenses and antibiotics. Bartonella henselae (B. henselae) is the causative agent of cat scratch disease, persistent bacteremia, and one of the most frequently reported causes of blood-culture negative endocarditis (BCNE) in patients. The ability of B. henselae to adhere to the heart valve, form a biofilm and vegetation to cause endocarditis increases the morbidity and mortality rate in infected patients. The presence of a trimeric autotransporter adhesin (TAA) called …
Culture-Based Analysis Of ‘Cave Silver’ Biofilms On Rocks In The Former Homestake Mine In South Dakota, Usa, Amanpreet K. Brar, David Bergmann
Culture-Based Analysis Of ‘Cave Silver’ Biofilms On Rocks In The Former Homestake Mine In South Dakota, Usa, Amanpreet K. Brar, David Bergmann
International Journal of Speleology
Tunnels in a warm, humid area of the 1478 m level of the Sanford Underground Research Facility (SURF), located in a former gold mine in South Dakota, USA, host irregular, thin whitish, iridescent biofilms, which appear superficially similar to ‘cave silver’ biofilms described from limestone and lava tube caves, despite the higher rock temperature (32°C) and differing rock surface (phyllite) present at SURF. In this study, we investigated the diversity of cultivable bacteria constituting the cave silver by using several media: CN agar, CN gellan gum and 0.1X R2A agar. The highest colony count (CFU/g of sample) was observed on …
Investigating The Proteinaceous Regulome Of The Acinetobacter Baumannii, Leila G. Casella
Investigating The Proteinaceous Regulome Of The Acinetobacter Baumannii, Leila G. Casella
USF Tampa Graduate Theses and Dissertations
Acinetobacter baumannii is an opportunistic pathogen that overtime has evolved into one of the most problematic pathogens due to its ability to overcome antibiotic pressures and harshly environments in the host and hospital environments. In this context, its genomic evolution due to its capacity to acquire genes that contribute to its pathogenic and antibiotic resistance nature has been the subject of research in the last decades providing with the identification of several proteins aiding in the process of pathogenicity. Although these findings have contributed to our understanding of A. baumannii pathogenic traits, the regulatory network that control their expression are …
Cyanobacterial And Algal Abundance And Biomass In Cave Biofilms And Relation To Environmental And Biofilm Parameters, Slađana Popović, Nataša Nikolić, Jelena Jovanović, Dragana Predojević, Ivana Trbojević, Ljiljana Manić, Gordana Subakov Simić
Cyanobacterial And Algal Abundance And Biomass In Cave Biofilms And Relation To Environmental And Biofilm Parameters, Slađana Popović, Nataša Nikolić, Jelena Jovanović, Dragana Predojević, Ivana Trbojević, Ljiljana Manić, Gordana Subakov Simić
International Journal of Speleology
Due to life in extreme environments, cyanobacteria and algae from cave biofilms that form at the entrances or deep inside the cave around artificial lights are of increasing interest to many scientists. It is well-known that many phototrophic microorganisms are first to colonize exposed substrata and produce the organic matter on which other biofilm constituents relly. Many studies dealing with phototrophic microorganisms from biofilms focus on the diversity and community composition of cyanobacteria and algae, while quantitative assessments are rarely implemented. Biofilm sampling was conducted in Degurić and Vernjikica Cave located in Western and Eastern Serbia, respectively. Ecological parameters (temperature, …
Cornus Officinalis Significantly Improves Oxidative Capacity And Promotes The Calcium-Dependent Transcription Factor, Nfatc2, In Human 1.1b4 Pancreatic Cell Line, Arielle Sharp, Alexis Coiner, Brant Burkhardt
Cornus Officinalis Significantly Improves Oxidative Capacity And Promotes The Calcium-Dependent Transcription Factor, Nfatc2, In Human 1.1b4 Pancreatic Cell Line, Arielle Sharp, Alexis Coiner, Brant Burkhardt
Molecular Biosciences Faculty Publications
Type 1 diabetes (T1D) is an autoimmune disease resulting in the destruction of pancreatic β cells (β-cells) and subsequent loss of insulin production. The only treatment for T1D is using exogenous insulin coupled with continual glucose monitoring following significant autoimmune destruction of β-cells. Novel interventional therapies are needed that can preserve and protect existing pancreatic β cells in individuals with early identified T1D autoimmunity. Our initial in-vitro evidence indicates Cornus officinalis (CO) may be able to serve in this function. What sets ethnopharmacology apart from conventional medicine is the simultaneous targeting of multiple mechanisms using a single herb due to …
Structural Basis Of Phosphatidylcholine Recognition By The C2–Domain Of Cytosolic Phospholipase A,2Α, Yoshinori Hirano, Yong-Guang Gao, Daniel J. Stephenson, Ngoc T. Vu, Lucy Malinina, Dhirendra K. Simanshu, Charles E. Chalfant, Dinshaw J. Patel, Rhoderick E. Brown
Structural Basis Of Phosphatidylcholine Recognition By The C2–Domain Of Cytosolic Phospholipase A,2Α, Yoshinori Hirano, Yong-Guang Gao, Daniel J. Stephenson, Ngoc T. Vu, Lucy Malinina, Dhirendra K. Simanshu, Charles E. Chalfant, Dinshaw J. Patel, Rhoderick E. Brown
Molecular Biosciences Faculty Publications
Ca2+-stimulated translocation of cytosolic phospholipase A2α (cPLA2α) to the Golgi induces arachidonic acid production, the rate-limiting step in pro-inflammatory eicosanoid synthesis. Structural insights into the cPLA2α preference for phosphatidylcholine (PC)-enriched membranes have remained elusive. Here, we report the structure of the cPLA2α C2-domain (at 2.2 Å resolution), which contains bound 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) and Ca2+ ions. Two Ca2+ are complexed at previously reported locations in the lipid-free C2-domain. One of these Ca2+ions, along with a third Ca2+, bridges the C2-domain to the DHPC phosphate group, which also interacts with Asn65. Tyr96 plays a key role in lipid …
Conserved Glycines Control Disorder And Function In The Cold-Regulated Protein, Cor15a, Oluwakemi T. Sowemimo, Patrick Knox-Brown, Wade M. Borcherds, Tobias Rindfleisch, Anja Thalhammer, Gary W. Daughdrill
Conserved Glycines Control Disorder And Function In The Cold-Regulated Protein, Cor15a, Oluwakemi T. Sowemimo, Patrick Knox-Brown, Wade M. Borcherds, Tobias Rindfleisch, Anja Thalhammer, Gary W. Daughdrill
Molecular Biosciences Faculty Publications
Cold-regulated (COR) 15A is an intrinsically disordered protein (IDP) from Arabidopsis thaliana important for freezing tolerance. During freezing-induced cellular dehydration, COR15A transitions from a disordered to mostly α-helical structure. We tested whether mutations that increase the helicity of COR15A also increase its protective function. Conserved glycine residues were identified and mutated to alanine. Nuclear magnetic resonance (NMR) spectroscopy was used to identify residue-specific changes in helicity for wildtype (WT) COR15A and the mutants. Circular dichroism (CD) spectroscopy was used to monitor the coil–helix transition in response to increasing concentrations of trifluoroethanol (TFE) and ethylene glycol. The impact of the COR15A …
Functional Inhibition Of Acid Sphingomyelinase Disrupts Infection By Intracellular Bacterial Pathogens, Chelsea L. Cockburn, Ryan S. Green, Sheela R. Damle, Rebecca K. Martin, Naomi N. Ghahrai, Punsiri M. Colonne, Marissa S. Fullerton, Daniel H. Conrad, Charles E. Chalfant, Daniel E. Voth, Elizabeth A. Rucks, Stacey D. Gilk, Jason A. Carlyon
Functional Inhibition Of Acid Sphingomyelinase Disrupts Infection By Intracellular Bacterial Pathogens, Chelsea L. Cockburn, Ryan S. Green, Sheela R. Damle, Rebecca K. Martin, Naomi N. Ghahrai, Punsiri M. Colonne, Marissa S. Fullerton, Daniel H. Conrad, Charles E. Chalfant, Daniel E. Voth, Elizabeth A. Rucks, Stacey D. Gilk, Jason A. Carlyon
Molecular Biosciences Faculty Publications
Intracellular bacteria that live in host cell–derived vacuoles are significant causes of human disease. Parasitism of low-density lipoprotein (LDL) cholesterol is essential for many vacuole-adapted bacteria. Acid sphingomyelinase (ASM) influences LDL cholesterol egress from the lysosome. Using functional inhibitors of ASM (FIASMAs), we show that ASM activity is key for infection cycles of vacuole-adapted bacteria that target cholesterol trafficking—Anaplasma phagocytophilum, Coxiella burnetii, Chlamydia trachomatis, and Chlamydia pneumoniae. Vacuole maturation, replication, and infectious progeny generation by A. phagocytophilum, which exclusively hijacks LDL cholesterol, are halted and C. burnetii, for which lysosomal cholesterol accumulation is bactericidal, …
Functional Analysis Of The Replication Fork Proteome Identifies Bet Proteins As Pcna Regulators, Sarah R. Wessel, Kareem N. Mohni, Jessica W. Luzwick, Huzefa Dungrawala, David Cortez
Functional Analysis Of The Replication Fork Proteome Identifies Bet Proteins As Pcna Regulators, Sarah R. Wessel, Kareem N. Mohni, Jessica W. Luzwick, Huzefa Dungrawala, David Cortez
Molecular Biosciences Faculty Publications
Identifying proteins that function at replication forks is essential to understanding DNA replication, chromatin assembly, and replication-coupled DNA repair mechanisms. Combining quantitative mass spectrometry in multiple cell types with stringent statistical cutoffs, we generated a high-confidence catalog of 593 proteins that are enriched at replication forks and nascent chromatin. Loss-of-function genetic analyses indicate that 85% yield phenotypes that are consistent with activities in DNA and chromatin replication or already have described functions in these processes. We illustrate the value of this resource by identifying activities of the BET family proteins BRD2, BRD3, and BRD4 in controlling DNA replication. These proteins …
P53 Phosphomimetics Preserve Transient Secondary Structure But Reduce Binding To Mdm2 And Mdmx, Robin Levy, Emily Gregory, Wade Borcherds, Gary W. Daughdrill
P53 Phosphomimetics Preserve Transient Secondary Structure But Reduce Binding To Mdm2 And Mdmx, Robin Levy, Emily Gregory, Wade Borcherds, Gary W. Daughdrill
Molecular Biosciences Faculty Publications
The disordered p53 transactivation domain (p53TAD) contains specific levels of transient helical secondary structure that are necessary for its binding to the negative regulators, mouse double minute 2 (Mdm2) and MdmX. The interactions of p53 with Mdm2 and MdmX are also modulated by posttranslational modifications (PTMs) of p53TAD including phosphorylation at S15, T18 and S20 that inhibits p53-Mdm2 binding. It is unclear whether the levels of transient secondary structure in p53TAD are changed by phosphorylation or other PTMs. We used phosphomimetic mutants to determine if adding a negative charge at positions 15 and 18 has any effect on the transient …