Physical Sciences and Mathematics Commons^™

Substrate Specificity In Abc Transporters Using The E. Coli Methionine Import System, John H. Guardado

Featured Student Work

ATP-binding cassette (ABC) transporters use the energy of ATP to move substrates across membranes against a concentration gradient. The role of ABC transporters is crucial in several essential cellular functions and mutations in ABC transporters in humans have been linked to several conditions, including cystic fibrosis, liver disease, and diabetes. Despite their central roles in homeostasis, the mechanism of ABC transporters remains poorly understood. Our research is focused on studying an ABC importer in E. coli, as a model system, to examine the mechanism of substrate specificity and transport. The bacterial methionine import system consists of a membrane-embedded transporter, MetNI, …

Usf Jamovi Tutorial Project: Open Education Resource, Aline Hitti, Saera Khan

USF OER Faculty Grant

Jamovi is an open source free software that USF staff, faculty and student can download to carry out any statistical analyses. The current report summarizes the progress made on an Open Education Resource Grant funded project, which aimed to created Jamovi tutorials. In this report, student feedback and faculty reaction are summarized after one semester of using the tutorials created.

Unboxing Cluster Heatmaps, Sophie J. Engle, S. Whalen, Alark Joshi, K. Pollard

Computer Science

Background: Cluster heatmaps are commonly used in biology and related fields to reveal hierarchical clusters in data matrices. This visualization technique has high data density and reveal clusters better than unordered heatmaps alone. However, cluster heatmaps have known issues making them both time consuming to use and prone to error. We hypothesize that visualization techniques without the rigid grid constraint of cluster heatmaps will perform better at clustering-related tasks.

Results: We developed an approach to “unbox” the heatmap values and embed them directly in the hierarchical clustering results, allowing us to use standard hierarchical visualization techniques as alternatives …

Altitudinal Shifts Of The Native And Introduced Flora Of California In The Context Of 20th-Century Warming, A. Wolf, Naupaka B. Zimmerman, W. R. Anderegg, P. E. Busby, J. Christensen

Biology Faculty Publications

Aim: The differential responses of plant species to climate change are of great interest and grave concern for scientists and conservationists. One underexploited resource for better understanding these changes are the records held by herbaria. Using these records to assess the responses of different groups of species across the entire flora of California, we sought to quantify the magnitude of species elevational shifts, to measure differences in shifts among functional groups and between native and introduced species, and to evaluate whether these shifts were related to the conservation of thermal niches.

Location: California.

Methods: To characterize these shifts in California, …

The Allosteric Regulatory Mechanism Of The Escherichia Coli Metni Methionine Atp Binding Cassette (Abc) Transporter, Janet G. Yang, D. C. Rees

Chemistry Faculty Publications

The MetNI methionine importer of Escherichia coli, an ATP binding cassette (ABC) transporter, uses the energy of ATP binding and hydrolysis to catalyze the high affinity uptake of d- and l-methionine. Early in vivo studies showed that the uptake of external methionine is repressed by the level of the internal methionine pool, a phenomenon termed transinhibition. Our understanding of the MetNI mechanism has thus far been limited to a series of crystal structures in an inward-facing conformation. To understand the molecular mechanism of transinhibition, we studied the kinetics of ATP hydrolysis using detergent-solubilized MetNI. We find that transinhibition is …

Using Field Data To Assess Model Predictions Of Surface And Ground Fuel Consumption By Wildfire In Coniferous Forests Of California, J M. Lydersen, B M. Collins, C M. Ewell, A L. Reiner, J A. Fites, C B. Dow, P Gonzalez, David Saah, J J. Battles

Environmental Science

Inventories of greenhouse gas (GHG) emissions from wildfire provide essential information to the state of California, USA, and other governments that have enacted emission reductions. Wildfires can release a substantial amount of GHGs and other compounds to the atmosphere, so recent increases in fire activity may be increasing GHG emissions. Quantifying wildfire emissions however can be difficult due to inherent variability in fuel loads and consumption and a lack of field data of fuel consumption by wildfire. We compare a unique set of fuel data collected immediately before and after six wildfires in coniferous forests of California to fuel consumption …

Lack Of Quantitative Training Among Early-Career Ecologists: A Survey Of The Problem And Potential Solutions, F. Barraquand, T. G. Ezard, P. Søgaard Jørgensen, Naupaka B. Zimmerman, S. Chamberlain, R. Salguero-Gómez, T. J. Curran, T. Poisot

Biology Faculty Publications

Proficiency in mathematics and statistics is essential to modern ecological science, yet few studies have assessed the level of quantitative training received by ecologists. To do so, we conducted an online survey. The 937 respondents were mostly early-career scientists who studied biology as undergraduates. We found a clear self-perceived lack of quantitative training: 75% were not satisfied with their understanding of mathematical models; 75% felt that the level of mathematics was “too low” in their ecology classes; 90% wanted more mathematics classes for ecologists; and 95% more statistics classes. Respondents thought that 30% of classes in ecology-related degrees should be …

Fire Behavior Modeling To Assess Net Benefits Of Forest Treatments On Fire Hazard Mitigation And Bioenergy Production In Northeastern California, David J. Ganz, David Saah, Klaus Barber, Mark Nechodom

Environmental Science

The fire behavior modeling described here, conducted as part of the Biomass to Energy (B2E) life cycle assessment, is funded by the California Energy Commission to evaluate the potential net benefits associated with treating and utilizing forest biomass. The B2E project facilitates economic, environmental, energy, and effectiveness assessments of the potential public benefits associated with: (1) various options for treatment, disposition, and utilization of forest biomass and (2) energy production from biomass produced by forest remediation activities. The study models forest conditions, fire behavior and fuel changes over a 40-year period, under three fuel treatment scenarios: no treatment; harvest and …

From Morphology To Neural Information: The Electric Sense Of The Skate, Marcelo Camperi, Timothy C. Tricas, Brandon R. Brown

Physics and Astronomy

Morphology typically enhances the fidelity of sensory systems. Sharks, skates, and rays have a well-developed electrosense that presents strikingly unique morphologies. Here, we model the dynamics of the peripheral electrosensory system of the skate, a dorsally flattened batoid, moving near an electric dipole source (e.g., a prey organism). We compute the coincident electric signals that develop across an array of the skate's electrosensors, using electrodynamics married to precise morphological measurements of sensor location, infrastructure, and vector projection. Our results demonstrate that skate morphology enhances electrosensory information. Not only could the skate locate prey using a simple population vector algorithm, but …

Two Purified Domains Of Telomerase Reverse Transcriptase Reconstitute Sequence-Specific Interactions With Rna, C. M. O'Connor, Cary K. Lai, Kathleen Collins

Biology Faculty Publications

Telomerase reverse transcriptase (TERT) and telomerase RNA (TER) function together to create a uniquely specialized polymerase. Here we have described for the first time domains of bacterially expressed Tetrahymena TERT that interacted directly with TER in the absence of assembly chaperones. We used quantitative binding assays to define TER sequence requirements for recognition by the high affinity RNA binding domain and an independent N-terminal RNA interaction domain. The TERT RNA binding domain and N-terminal RNA interaction domain had distinct, nonoverlapping requirements for TER sequence and structure that together accounted for all of the sites of TER contact inferred for full-length …

Modeling An Electrosensory Landscape: Behavioral And Morphological Optimization In Elasmobranch Prey Capture, Brandon R. Brown

Physics and Astronomy

Most biological sensory systems benefit from multiple sensors. Elasmobranchs (sharks, skates and rays) possess an array of electroreceptive organs that facilitate prey location, mate location and navigation. Here, the perceived electrosensory landscape for an elasmobranch approaching prey is mathematically modeled. The voltages that develop simultaneously in dozens of separate sensing organs are calculated using electrodynamics. These voltages lead directly to firing rate modifications in the primary afferent nerves. The canals connecting the sense organs to an elasmobranch's surface exhibit great variation of location and orientation. Here, the voltages arising in the sense organs are found to depend strongly on the …

Characterization Of Chimeric Lipopolysaccharides From Escherichia Coli Strain Jm109 Transformed With Lipooligosaccharide Synthesis Genes (Lsg) From Haemophilus Influenzae, Nancy J. Phillips, T. J. Miller, J. J. Engstrom, William Melaugh, R. Mclaughlin, M A. Apicella, B W. Gibson

Chemistry Faculty Publications

Previously, we reported the expression of chimeric lipopolysaccharides (LPS) in Escherichia coli strain JM109 (a K-12 strain) transformed with plasmids containing Haemophilus influenzae lipooligosaccharide synthesis genes (lsg) (Abu Kwaik, Y., McLaughlin, R. E., Apicella, M. A., and Spinola, S. M. (1991) Mol. Microbiol. 5, 2475–2480). In this current study, we have analyzed the O-deacylated LPS and free oligosaccharides from three transformants (designated pGEMLOS-4, pGEMLOS- 5, and pGEMLOS-7) by matrix-assisted laser desorption ionization, electrospray ionization, and tandem mass spectrometry techniques, along with composition and linkage analyses. These data show that the chimeric LPS consist of the complete E. coli LPS core …

Relationship Between Udp-Glucose 4-Epimerase Activity And Oligoglucose Glycoforms In Two Strains Of Neisseria Meningitidis, F K. Lee, B W. Gibson, William Melaugh, A Zaleski, M A. Apicella

Chemistry Faculty Publications

Sodium dodecyl sulfate-polyacrylamide gel analysis of lipooligosaccharide (LOS) from Neisseria meningitidis has demonstrated considerable microheterogeneity in the variable region of LOS due to the presence of novel glycoforms. As a step toward understanding the basis for the expression of these novel glycoforms, we have examined the LOS structures and UDP-glucose 4-epimerase (epimerase) activity levels in two strains (NMB and MA-1) and their respective galE mutants. Strain NMB was found to have low epimerase activity and to contain multiple glycoforms, some of which appear to contain only glucose sugars. The galE mutant had only the oligoglucose glycoforms. Strain MA-1 had higher …

Identification Of The Adp-L-Glycero-D-Manno-Heptose-6-Epimerase (Rfad) And Heptosyltransferase Ii (Rfaf) Biosynthesis Genes From Nontypeable Haemophilus Influenzae 2019, W A. Nichols, B W. Gibson, William Melaugh, N G. Lee, M Sunshine, M A. Apicella

Chemistry Faculty Publications

Haemophilus influenzae is an important human pathogen. The lipooligosaccharide (LOS) of H. influenzae has been implicated as a virulence determinant. To better understand the assembly of LOS in nontypeable H. influenzae (NtHi), we have cloned and characterized the rfaD and rfaF genes of NtHi 2019, which encode the ADP-L-glycero-D-manno-heptose-6-epimerase and heptosyltransferase II enzymes, respectively. This cloning was accomplished by the complementation of Salmonella typhimurium lipopolysaccharide (LPS) biosynthesis gene mutants. These deep rough mutants are novobiocin susceptible until complemented with the appropriate gene. In this manner, we are able to use novobiocin resistance to select for specific NtHi LOS inner core …

Characterization Of A Transposon Tn916-Generated Mutant Of Haemophilus Ducreyi 35000 Defective In Lipooligosaccharide Biosynthesis, B W. Gibson, A A. Campagnari, William Melaugh, M A. Apicella, S Grass, Jing Wang, Katherine L. Palmer, R S. Munson

Chemistry Faculty Publications

To define the role of the surface lipooligosaccharide (LOS) of Haemophilus ducreyi in the pathogenesis of chancroid, Tn916 mutants of H. ducreyi 35000 defective in expression of the murine monoclonal antibody (MAb) 3F11 epitope on H. ducreyi LOS were identified by immunologic screening. One mutant, designated 1381, has an LOS which lacks the MAb 3F11 epitope and migrates with an increased mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The gene disrupted by the Tn916 element in strain 1381 was identified by cloning the sequences flanking the Tn916 element. The sequences were then used to probe a lambda DASHII genomic library. …

The Lipooligosaccharides Of Haemophilus Ducreyi Are Highly Sialylated, William Melaugh, A A. Campagnari, B W. Gibson

Chemistry Faculty Publications

The major lipooligosaccharides of the sexually transmitted pathogen Haemophilus ducreyi 35000 have been previously found to terminate in N-acetyllactosamine and sialyl-N-acetyllactosamine, Neu5Ac alpha 2-->3Gal beta 1-->4GlcNAc (W. Melaugh, N. J. Phillips, A. A. Campagnari, M. V. Tullius, and B. W. Gibson, Biochemistry 33: 13070-13078, 1994). In this study, mass spectrometry and composition analyses have shown that the lipooligosaccharides from three other H. ducreyi strains also contain N-acetyllactosamine and are highly sialylated (approximately 30 to 50%), although one African strain was found to contain neither of these structural features.

Use Of Pyocin To Select A Haemophilus Ducreyi Variant Defective In Lipooligosaccharide Biosynthesis, A A. Campagnari, R Karalus, M A. Apicella, William Melaugh, A J. Lesse, B W. Gibson

Chemistry Faculty Publications

Haemophilus ducreyi, a cause of genital ulcer disease in developing countries, appears to facilitate the heterosexual transmission of the human immunodeficiency virus in Africa. Despite an increase in studies of this gram-negative human pathogen, little is known about the pathogenesis of chancroid. Our studies have shown that the lipooligosaccharides (LOS) of H. ducreyi may play an important role in ulcer formation. Monoclonal antibody and mass spectrometric analyses identified a terminal trisaccharide present on H. ducreyi LOS that is immunochemically similar to human paragloboside. This epitope is present on the LOS of Neisseria gonorrhoeae, and it may be the site of …

Considerations For Evaluating Ultraviolet Radiation-Induced Genetic Damage Relative To Antarctic Ozone Depletion, Deneb Karentz

Biology Faculty Publications

Springtime ozone depletion over the Antarctic results in increased UVB in local marine environments. It has been established that decreases in primary productivity occur with decreases in ozone concentrations, but the impact of increased UVB on the functioning and stability of the ecosystem has not yet been determined. Very little has been done to evaluate the potential for genetic damage caused by the increase in UVB, and this type of damage is most significant relative to the fitness and maintenance of populations. An essential problem in evaluating genotoxic effects is the lack of appropriate techniques to sample and quantify genetic …

Investigation Of The Structural Heterogeneity Of Lipooligosaccharides From Pathogenic Haemophilus And Neisseria Species And Of R-Type Lipopolysaccharides From Salmonella Typhimurium By Electrospray Mass Spectrometry, B W. Gibson, William Melaugh, Nancy J. Phillips, M A. Apicella, A A. Campagnari, J M. Griffiss

Chemistry Faculty Publications

Heterogeneity in the lipooligosaccharides (LOS) of pathogenic Haemophilus and Neisseria species is evident from the multiplicity of components observed with electrophoretic analyses. Knowledge of the precise structures that make up these diverse LOS molecules is clearly the key to reaching an understanding of pathogenic processes such as phase variation and molecular mimicry. Except for a few cases, little is known about the specific structural features of LOS that underlie phase variation and molecular mimicry, partly because of the inherent difficulties in the structural elucidation of these complex glycolipids. In the lipopolysaccharides (LPS) from Salmonella typhimurium and Escherichia coli, rough, or …

Partial Characterization Of The Major Lipooligosaccharide From A Strain Of Haemophilus Ducreyi, The Causative Agent Of Chancroid, A Genital Ulcer Disease, William Melaugh, Nancy J. Phillips, A A. Campagnari, R Karalus, B W. Gibson

Chemistry Faculty Publications

The first preliminary structure of a surface lipooligosaccharide from Haemophilus ducreyi has been determined. The major oligosaccharide was released by mild acid hydrolysis and analyzed by liquid secondary ion and tandem mass spectrometry. The mass spectral data combined with composition and methylation analysis yielded the most probable structure; Gal1----4GlcNAc1----3Gal1----4Hep1----6Glc1----( Hep1----2Hep1----)3,4Hep1---- KDO, where the reducing terminal 3-deoxy-D-manno-octulosonic acid (or KDO) exists in an anhydro form. This anhydro species results from the elimination of a phosphate from C-4 of KDO during mild acid hydrolysis. The core heptose trisaccharide consists of L-glycero-D-manno-heptose, but analysis of the peracetylated sugars indicated that the 1,4-linked heptose …