Life Sciences Commons^™

Multiple Factors Independently Regulate Hila And Invasion Gene Expression In Salmonella Enterica Serovar Typhimurium, Robin L. Lucas, C. Phoebe Lostroh, Concetta C. Dirusso, Michael P. Spector, Barry L. Wanner, Catherine A. Lee

Department of Biochemistry: Faculty Publications

HilA activates the expression of Salmonella enterica serovar Typhimurium invasion genes. To learn more

about regulation of hilA, we isolated Tn5 mutants exhibiting reduced hilA and/or invasion gene expression. In

addition to expected mutations, we identified Tn5 insertions in pstS, fadD, flhD, flhC, and fliA. Analysis of the

pstS mutant indicates that hilA and invasion genes are repressed by the response regulator PhoB in the absence

of the Pst high-affinity inorganic phosphate uptake system. This system is required for negative control of the

PhoR-PhoB two-component regulatory system, suggesting that hilA expression may …

The Holliday Junction In An Inverted Repeat Dna Sequence: Sequence Effects On The Structure Of Four-Way Junctions, Brandt F. Eichman, Jeff Vargason, Blaine H.M. Mooers, P. Shing Ho

Faculty Publications - Department of Biological & Molecular Science

Holliday junctions are important structural intermediates in recombination, viral integration, and DNA repair. We present here the single-crystal structure of the inverted repeat sequence d(CCGGTACCGG) as a Holliday junction at the nominal resolution of 2.1 Å. Unlike the previous crystal structures, this DNA junction has B-DNA arms with all standard Watson–Crick base pairs; it therefore represents the intermediate proposed by Holliday as being involved in homologous recombination. The junction is in the stacked-X conformation, with two interconnected duplexes formed by coaxially stacked arms, and is crossed at an angle of 41.4° as a right-handed X. A sequence comparison with previous …

Job Compensation In The Biotechnology Core Laboratory, Debra A. Mcmillen, William M. Old, Janice L. Bleibaum, Charles M. Nicolet, Katheryn R. Resing, Gautam Sarath, Satya P. Yadav

Gautam Sarath Publications

The results of a poll of core facility professionals by the Association of Biomolecular Resource Facilities (ABRF) Survey Committee.

Two-Step Processing Of Human Frataxin By Mitochondrial Processing Peptidase, Patrizia Cavadini, Jiri Adamec, Franco Taroni, Oleksandr Gakh, Grazia Isaya

Department of Biochemistry: Faculty Publications

We showed previously that maturation of the human

frataxin precursor (p-fxn) involves two cleavages by the

mitochondrial processing peptidase (MPP). This observation

was not confirmed by another group, however,

who reported only one cleavage. Here, we demonstrate

conclusively that MPP cleaves p-fxn in two sequential

steps, yielding a 18,826-Da intermediate (i-fxn) and a

17,255-Da mature (m-fxn) form, the latter corresponding

to endogenous frataxin in human tissues. The two cleavages

occur between residues 41–42 and 55–56, and both

match the MPP consensus sequence RX ↓ (X/S). Recombinant

rat and yeast MPP catalyze the pài step 4 and 40 …

Rbcs Suppressor Mutations Improve The Thermal Stability And Co2/O2 Specificity Of Rbcl-Mutant Ribulose-1,5-Bisphosphate Carboxylaseyoxygenase, Yu-Chan Du, Seokjoo Hong, Robert J. Spreitzer

Department of Biochemistry: Faculty Publications

In the green alga Chlamydomonas reinhardtii, a Leu²⁹⁰-to-Phe (L²⁹⁰F) substitution in the large subunit of ribulose-1,5-bisphosphate carboxylaseyoxygenase (Rubisco), which is coded by the chloroplast rbcL gene, was previously found to be suppressed by second-site Ala222-to-Thr and Val²⁶²-to-Leu substitutions. These substitutions complement the photosynthesis deficiency of the L290F mutant by restoring the decreased thermal stability, catalytic efficiency, and CO₂/O₂ specificity of the mutant enzyme back to wild-type values. Because residues 222, 262, and 290 interact with the loop between β strands A and B of the Rubisco small subunit, which is …

Applications Of The Atomic Force Microscope Within The Fields Of Haematology And Cytogenetics, Margaret O'Reilly-Crowley

Theses

Atomic force microscopy [AFM] has become a rapidly developing interdisciplinary field of research in recent years. The technique, including instrumentation and specimen preparation methods, is now more sophisticated and its application has increased dramatically. The key advantage of AFM is in providing three-dimensional images of samples imaged. Both size and shape of samples imaged can be obtained allowing their subsequent quantification. Red blood cells [RBCs] and chromosomes were imaged in this work. A brief overview of the current status of biomedical AFM, has been presented in the review.

Measurement of mean cell volume [MCV] of immobilised red blood cells with …

The Specific Immune Response In Rainbow Trout: Somatic Hypermutation And Vh Gene Utilization, Teresa D. Lewis

Dissertations, Theses, and Masters Projects

The study of antibody responses in prominent aquaculture species such as the rainbow trout, Oncorhynchus mykiss, can facilitate vaccine development and contribute to producing useful paradigms of adaptive immunity in lower vertebrates. Thus, it is essential to identify genes responsible for antibody responses. In the mouse model, hybridoma technology allows for the association of monoclonal antibodies possessing various affinities for antigen with specific VH sequences, gene family utilization, and other molecular events (i.e. somatic hypermutation) that occur during the specific immune response. The absence of a comparable hybridoma technology in piscine systems has limited similar studies of fish immunogenetics to …

Function And Regulation Of Alternative Isoforms Of The Transcription Factor Pax-5 During B Lymphocyte Differentiation, Marina A. Lowen

Dissertations, Theses, and Masters Projects

No abstract provided.

Biochemical Composition Of Particles And Dissolved Organic Matter Slong An Estuarine Gradient: Sources And Implications For Dom Reactivity, Antonio Mannino, H. Rodger Harvey

OES Faculty Publications

The chemical composition of high molecular weight dissolved organic matter (DOM) and particulate organic matter (POM) was examined along the salinity gradient of the Delaware Estuary. DOM was collected and fractionated by tangential-flow ultrafiltration into 1-30 kDa (HDOM; high molecular weight) and 30 kDa to 0.2 μm (VHDOM; very high molecular weight) and compared to particles collected in parallel. Polysaccharides comprised 12-43% of particulate organic carbon (POC), 30-56% of VHDOM carbon, and 7.5-19% of HDOM carbon. Hydrolyzable amino acids comprised 17-38% of POC, 5.4-12% of VHDOM carbon, and 1.5-4.2% of HDOM carbon. Only 7-43% of dissolved organic nitrogen in VHDOM …

Human Gonadotropin-Releasing Hormone (Hgnrh) Gene Expression And Hormone Regulation In Human Placental Jeg-3 Cells, Heming Zheng

Theses and Dissertations in Biomedical Sciences

Using the human placental choriocarcinoma JEG-3 cell line as an in vitro human placental model, I studied the mechanisms of the tissue-specific expression and steroid hormone regulation of the hGnRH gene in the human placenta. The results showed that all of the previously identified four elements are required for the full activity of the hGnRH upstream promoter in JEG-3 cells, while the element 4 (FP4, −987/−968) is the most important. Studies performed with 5′ end deletion of this region confirmed these observations. Further, supershift assay using Oct-1 antibody demonstrated the involvement of Oct-1 in the FP4 DNA-protein interaction in JEG-3 …

Detection And Modeling Of Radiation Induced Effects In Tissues By Dielectric Spectroscopy, Dorin A. Todor

Physics Theses & Dissertations

The work presented here is applied physics research in the field of radiation treatment. We address the development of a new and innovative method, in vivo and possibly non-invasive, for tumor and healthy tissues control during and after the radiation treatment. The radiation treatment is delivered in an almost standardized manner for particular classes of tumors. The large variance in the individual radio sensitivity of healthy tissues and tumors often leads to local recurrence of neoplastic growth and/or distant metastatic disease which often remains untreated. The method is based on the measurement and analysis of electrical impedance data in …

Cytotoxic T Cell Response To Influenza Vaccination In Older Adults, Caroline R. Letter

Chemistry & Biochemistry Theses & Dissertations

Influenza infections result in activation of cellular and humoral immune responses, leading to stimulation of cytotoxic T cells (CTL) and helper T cells with subsequent viral clearance.

By using an assay of granzyme B activity, the in vivo cellular-mediated immune response of old adults was found to be significantly decreased in comparison to young adults. The effect of aging on the CTL response was further investigated via enzyme-linked immunospot technique. A significant increase in number of virus-specific T cells occurred in older adults after vaccination, although the magnitude of the CTL response decreased in comparison to young adults. These data …

Compartmental Specificity Of Cellular Membrane Fusion Encoded In Snare Proteins, James Mcnew, Frank Parlati, Ryorichi Fukuda, Robert Johnston, Keren Paz, Fabienne Paumet, Thomas Sollner, James Rothman

Fabienne Paumet

No abstract provided.

Dependence Of The Extent Of Ion Association With Polynucleotides And Peptides As A Function Of Solution Dielectric., David Mascotti, A. Salwan

David P. Mascotti

No abstract provided.

Green Fluorescent Protein As A Quantitative Reporter Of Relative Promoter Activity In E-Coli., James Lissemore, J. Jankowski, C. Thomas, David Mascotti, P. Dehaseth

David P. Mascotti

No abstract provided.

Soluble Nsf Attachment Protein Receptors (Snares) In Rbl-2h3 Mast Cells: Functional Role Of Syntaxin4 In Exocytosis And Identification Of A Vamp8-Containing Secretory Compartment, Fabienne Paumet, Joelle Le Mao, Sophie Martin, Thierry Galli, Bernard David, Ulrich Blank, Michele Roa

Fabienne Paumet

No abstract provided.

Electrogenic Sodium–Sodium Exchange Carried Out By Na,K-Atpase Containing The Amino Acid Substitution Glu779ala, José Argüello, R.Daniel Peluffo, Jerry Lingrel, Joshua Berlin

José M. Argüello

Na,K-ATPase containing the amino acid substitution glutamate to alanine at position 779 of the a subunit (Glu779Ala) supports a high level of Na-ATPase and electrogenic Na1–Na1 exchange activityin the absence of K1. In microsomal preparations of Glu779Ala enzyme, the Na1 concentration for half maximal activation of Na-ATPase activity was 161 6 14 mM (n 5 3). Furthermore, enzyme activity with 800 mM Na1 was found to be similar in the presence and absence of 20 mM K1. These results showed that Na1, with low affinity, could stimulate enzyme turnover as effectively as K1. To gain further insight into the mechanism …

The Role Of Na,K-Atpase Alpha Subunit Serine 775 And Glutamate 779 In Determining The Extracellular K+ And Membrane Potential-Dependent Properties Of The Na,K-Pump, José Argüello, R.Daniel Peluffo, Joshua Berlin

José M. Argüello

The roles of Ser775 and Glu779, two amino acids in the putative fifth transmembrane segment of the Na,K-ATPase a subunit, in determining the voltage and extracellular K1 (K1o) dependence of enzyme-mediated ion transport, were examined in this study. HeLa cells expressing the a1 subunit of sheep Na,K-ATPase were voltage clamped via patch electrodes containing solutions with 115 mM Na1 (378C). Na,K-pump current produced by the ouabain-resistant control enzyme (RD), containing amino acid substitutions Gln111Arg and Asn122Asp, displayed a membrane potential and K1o dependence similar to wild-type Na,K-ATPase during superfusion with 0 and 148 mM Na1-containing salt solutions. Additional substitution of …

N-Methyl-D-Aspartate Receptor Subunit Nr2b Is Widely Expressed Throughout The Rat Diencephalon: An Immunohistochemical Study, Arshad Khan, B. Glenn Stanley, Lisa Bozzetti, Christina Chin, Cyndi Stivers, Margarita Curras-Collazo

Arshad M. Khan, Ph.D.

No abstract provided.