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Molecular Biology

1992

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Articles 1 - 28 of 28

Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Circular Dichroism And Molecular Modeling Yield A Structure For The Complex Of Human Immunodeficiency Virus Type 1 Trans-Activation Response Rna And The Binding Region Of Tat, The Trans-Acting Transcriptional Activator, Erwann P. Loret, Philippe T. Georgel, W. Curtis Johnson Jr., Pui Shing Ho Oct 1992

Circular Dichroism And Molecular Modeling Yield A Structure For The Complex Of Human Immunodeficiency Virus Type 1 Trans-Activation Response Rna And The Binding Region Of Tat, The Trans-Acting Transcriptional Activator, Erwann P. Loret, Philippe T. Georgel, W. Curtis Johnson Jr., Pui Shing Ho

Biological Sciences Faculty Research

Transcription in the human immunodeficiency virus type 1 (HIV-1) retrovirus is regulated by binding the viral Tat protein (trans-acting transcriptional activator) to the trans-activation response (TAR) RNA sequence. Here, vacuum UV circular dichroism (VUV-CD) is used to study the structure of TAR and its complex with two peptide fragments that are important for Tat binding to TAR. The VUV-CD spectrum of TAR is typical of A-form RNA and is minimally perturbed when bound to either the short or the long Tat peptide. The CD spectra ofthe complexes indicate an extended structure in the argnine-rich region of Tat from amino acid ...


The Cpce And Cpcf Genes Of Synechococcus Sp. Pcc 7002. Construction And Phenotypic Characterization Of Interposon Mutants, J Zhou, G E. Gasparich, V L. Stirewalt, R De Lorimier, D A. Bryant Aug 1992

The Cpce And Cpcf Genes Of Synechococcus Sp. Pcc 7002. Construction And Phenotypic Characterization Of Interposon Mutants, J Zhou, G E. Gasparich, V L. Stirewalt, R De Lorimier, D A. Bryant

Gail Gasparich

The 3' region of the cpc operon of Synechococcus sp. PCC 7002 has been sequenced, transcriptionally characterized, and analyzed by interposon mutagenesis. The cpc operon contains six genes, 5' cpcB-cpcA-cpcC-cpcD-cpcE-cpcF 3', and gives rise to at least eight (more likely ten) discrete mRNA transcripts. The steady-state levels of transcripts for the cpcE and cpcF genes are very low and are estimated to represent only about 1-2% of the total transcripts arising from the cpc locus. The cpcE gene predicts a protein of 268 amino acid residues, whereas the cpcF gene predicts a protein of 205 amino acid residues. The deduced ...


Saccharomyces Cerevisiae Sec59 Cells Are Deficient In Dolichol Kinase Activity, Loree Heller, Peter Orlean, W. Lee Adair Jr. Aug 1992

Saccharomyces Cerevisiae Sec59 Cells Are Deficient In Dolichol Kinase Activity, Loree Heller, Peter Orlean, W. Lee Adair Jr.

Bioelectrics Publications

The temperature-sensitive Saccharomyces cerevisiae mutant sec59 accumulates inactive and incompletely glycosylated protein precursors in its endoplasmic reticulum at the restrictive temperature. O-mannosylation and glycosyl phosphatidylinositol membrane anchoring of protein are also abolished, consistent with a deficiency in dolichyl phosphate mannose. Membranes prepared from sec59 cells that had been shifted to the restrictive temperature, however, made normal amounts of dolichyl phosphate mannose when exogenous dolichyl phosphate was supplied, but dolichyl phosphate mannose synthesis was severely depressed in the absence of exogenous dolichyl phosphate. Quantitative measurements of dolichyl phosphate in sec59 cells showed that the levels were decreased to 48% of wild ...


Isolation And Characterization Of The Triiodothyronine Regulatory Region Of The Fatty Acid Synthase Gene In Avian Species, Delfina Rivera Valencia Aug 1992

Isolation And Characterization Of The Triiodothyronine Regulatory Region Of The Fatty Acid Synthase Gene In Avian Species, Delfina Rivera Valencia

Master's Theses

Fatty acids are utilized for energy storage in the form of triglycerides or neutral fats. One way to regulate fatty acid biosynthesis is to regulate the activity of fatty acid synthase (FAS), the enzyme that catalyzes the reaction. Previous studies have shown that regulation of FAS activity and synthesis is regulated by the hormone triiodothyronine, T3, at the level of transcription. Thus, isolation and characterization of the promoter region o f the FAS gene will enable the identification of the thyroid hormone response elements (TREs). This study could shed light on the mechanisms on how this gene can be ...


Distinct Permissive Pathways Mediate The Effects Of Nerve Growth Factor And Lithium On Neurotensin/Neuromedin N Gene Expression In Pc12 Cells: A Thesis, Bryant Paul Bullock Jun 1992

Distinct Permissive Pathways Mediate The Effects Of Nerve Growth Factor And Lithium On Neurotensin/Neuromedin N Gene Expression In Pc12 Cells: A Thesis, Bryant Paul Bullock

GSBS Dissertations and Theses

This thesis examines the effects of nerve growth factor (NGF) and lithium on the regulation of neurotensin/neuromedin N (NT/N) gene expression in PC12 pheochromocytoma cells. In PC12 cells, the expression of the rat NT/N gene is strictly dependent on simultaneous exposure to combinations of NGF, glucocorticoids, activators of adenylate cyclase, and lithium. Transient transfection experiments indicated that a consensus AP-1 site located within the NT/N promoter is the principal target of NGF and lithium action. NGF rapidly, but transiently, induces the expression of several AP-1 genes in PC12 cells, suggesting that the effect of NGF on ...


Protein Translocation Across The Er Requires A Functional Gtp Binding Site In The Alpha Subunit Of The Signal Recognition Particle Receptor, Peter J. Rapiejko, Reid Gilmore May 1992

Protein Translocation Across The Er Requires A Functional Gtp Binding Site In The Alpha Subunit Of The Signal Recognition Particle Receptor, Peter J. Rapiejko, Reid Gilmore

Open Access Articles

The signal recognition particle (SRP)-mediated translocation of proteins across the RER is a GTP dependent process. Analysis of the primary amino acid sequence of one protein subunit of SRP (SRP54), as well as the alpha subunit of the SRP receptor (SR alpha), has indicated that these proteins contain predicted GTP binding sites. Several point mutations confined to the GTP binding consensus elements of SR alpha were constructed by site specific mutagenesis to define a role for the GTP binding site in SR alpha during protein translocation. The SR alpha mutants were analyzed using an in vitro system wherein SR ...


Enzyme Cytochemical Localization Of Na,K-Atpase And Immunohistochemical Localization Of Vinculin In Frog And Mouse Liver Tissue, Douglas Mcinnis Apr 1992

Enzyme Cytochemical Localization Of Na,K-Atpase And Immunohistochemical Localization Of Vinculin In Frog And Mouse Liver Tissue, Douglas Mcinnis

Life and Environmental Sciences Undergraduate Theses

Means of localizing molecules are essential in contemporary biological research attempting to unlock the mysteries of how cells function, interact, and go wrong. Enzyme cytochemistry and immunohistochemistry are two methods which utilize different characteristics of given molecules to localize them in tissue. The objective of this study was to localize two proteins, Na,K-ATPase and vinculin, in frog and mouse liver sections using these methods. Na,KATPase activity contributes to overall p-nitrophenylphosphatase (jpNPPase) activity, and sites of p-NPPase activity were determined using an enzyme cytochemical method. Sites of p-NPPase activity were thus extrapolated to indicate regions of probable Na,K-ATPase ...


Virucidal Activities Of Amphipathic Compounds Against Lipid-Containing Bacteriophages, Gregory R. Keyock Jan 1992

Virucidal Activities Of Amphipathic Compounds Against Lipid-Containing Bacteriophages, Gregory R. Keyock

Theses and Dissertations

No abstract provided.


The Cγ Subunit Is A Unique Isozyme Of The Camp-Dependent Protein Kinase, Stephen J. Beebe, Paul Salomonsky, Tore Jahnsen, Yixin Li Jan 1992

The Cγ Subunit Is A Unique Isozyme Of The Camp-Dependent Protein Kinase, Stephen J. Beebe, Paul Salomonsky, Tore Jahnsen, Yixin Li

Bioelectrics Publications

There are at least three isozymes (Cα, Cβ, and Cγ) of the mammalian catalytic (C) subunit of cAMP-dependent protein kinase (PKA) (Beebe, S., Oyen, O., Sandberg, M., Froysa, A., Hansson, V., and Jahnsen, T. (1990) Mol. Endocrinol. 4, 465-475). To compare the Cγ and Cα isozymes, the respective cDNAs were expressed in permanently transformed Kin-8 PKA-deficient Y1 adrenal cells using the mouse metallothionein promoter. The recombinant C subunits were characterized as immunoreactive, zinc-inducible, cAMP-dependent kinase activities. In contrast to Cα, histone was a better substrate than Leu-Arg-Arg-Ala-Ser-Leu-Gly (Kemptide) for Cγ. Furthermore, Cγ histone kinase activity was not inhibited by the ...


Association Of Tetrapyrrole Intermediates In The Bacteriochlorophyll A Biosynthetic Pathway With The Major Outer-Membrane Porin Protein Of Rhodobacter Capsulatus, David Bollivar Jan 1992

Association Of Tetrapyrrole Intermediates In The Bacteriochlorophyll A Biosynthetic Pathway With The Major Outer-Membrane Porin Protein Of Rhodobacter Capsulatus, David Bollivar

Scholarship

No abstract provided.


Genetic And Biochemical Characterization Of The P21[Superscript Ras]-Related Protein Rho1p In The Yeast Saccharomyces Cerevisiae , Joni Marie Seeling Johnson Jan 1992

Genetic And Biochemical Characterization Of The P21[Superscript Ras]-Related Protein Rho1p In The Yeast Saccharomyces Cerevisiae , Joni Marie Seeling Johnson

Retrospective Theses and Dissertations

Rho1p is a guanine nucleotide binding protein coded for by the Saccharomyces cerevisiae gene RHO1, which is homologous to p21[superscript] ras and other members of the ras superfamily of small molecular weight GTP binding proteins. This protein family is highly conserved in evolution and in some instances is known to be involved in control of cell proliferation. The precise physiological functions of most p21[superscript] ras-related proteins, however, are not known. In order to gain further insight into the physiological function of rho proteins, this research addressed the genetic and biochemical characterization of Rho1p, as well as the subcellular ...


Fluorescence Studies And Consequences Of Amino Acid Substitutions With Escherichia Coli Adenylosuccinate Synthetase , Chandrasen Soans Jan 1992

Fluorescence Studies And Consequences Of Amino Acid Substitutions With Escherichia Coli Adenylosuccinate Synthetase , Chandrasen Soans

Retrospective Theses and Dissertations

Probes were introduced into the active site of adenylosuccinate synthetase from Escherichia coli for fluorescence energy transfer measurements and to measure enzyme conformational changes attending ligand binding. The distances from fluorescent reporter groups covalently bound to a highly reactive cysteinyl residue (Cys[superscript]291) of adenylosuccinate synthetase, to active site binding ligands were determined. Energy transfer from formycin 5[superscript]'-monophosphate, a competitive inhibitor of IMP, to Cys[superscript]291, modified with the fluorescent chromophore N-(iodoacetylaminoethyl)-5-naphthylamine-1-sulfonate, indicated that the chromophores are 17 A apart in the presence or absence of GTP. Analogous experiments using tetramethylrhodamine maleimide to modify ...


Characterization Of The Saccharomyces Cerevisiae Gene Mgp1, A Novel Member Of The Ras Gene Superfamily That Functions In The Mitochondrial Biogenesis , Shu-Hwa Lee Tzeng Jan 1992

Characterization Of The Saccharomyces Cerevisiae Gene Mgp1, A Novel Member Of The Ras Gene Superfamily That Functions In The Mitochondrial Biogenesis , Shu-Hwa Lee Tzeng

Retrospective Theses and Dissertations

This dissertation characterizes Mgp1p, a mitochondrial protein that functions in mitochondrial biogenesis. Mgp1p is the first protein related to the p21[superscript] H-ras oncoprotein shown to function in mitochondria;The Saccharomyces cerevisiae gene MGP1 was isolated based on its ability to restore respiratory competence to a strain containing the nuclear petite mutation msg1-1. MGP1 codes for a protein of approximately 58 kDa, with an amino terminal sequence typical of a mitochondrial targeting peptide. A 190 residue region of Mgp1p shares approximately 30% amino acid sequence identity with any member of p21[superscript] ras-related protein superfamily, with particularly strong homology in ...


Deletion Analysis Of The Starch Binding Domain From Aspergillus Glucoamylase , Luojing Chen Jan 1992

Deletion Analysis Of The Starch Binding Domain From Aspergillus Glucoamylase , Luojing Chen

Retrospective Theses and Dissertations

Glucoamylase (GA) from Aspergillus (EC 3.2.1.3) exists in two forms: GAI (amino acids 1-616) and GAII (amino acids 1-512). GAII is identical to GAI but lacks the C-terminal region (amino acids 513-616). While both forms of glucoamylase are able to digest soluble starch, only GAI has the ability to bind to and therefore hydrolyze native starch granules. To further examine the location of the starch binding domain at the C-terminus of GAI and to determine how many amino acid residues of GAI are required for starch binding, six [beta]-galactosidase fusion proteins were constructed. Affinity purified fusion ...


Effects Of Temperature And Ethanol On Maturation And Germination Of Trichoderma Reesei, Kum Kum Sharma Jan 1992

Effects Of Temperature And Ethanol On Maturation And Germination Of Trichoderma Reesei, Kum Kum Sharma

Theses and Dissertations

No abstract provided.


Nucleotide Sequence Of S-Adenosyl-L-Methionine: Magnesium Protoporphyrin Methyltransferase From Rhodobacter Capsulatus, David Bollivar Jan 1992

Nucleotide Sequence Of S-Adenosyl-L-Methionine: Magnesium Protoporphyrin Methyltransferase From Rhodobacter Capsulatus, David Bollivar

Scholarship

No abstract provided.


Cytophotometric Estimation Of Nuclear Dna Content Variation In Ten Species Of Geniculate Coralline Algae (Corallinaceae, Rhodophyta), Jeffrey Craig Bailey Jan 1992

Cytophotometric Estimation Of Nuclear Dna Content Variation In Ten Species Of Geniculate Coralline Algae (Corallinaceae, Rhodophyta), Jeffrey Craig Bailey

Dissertations, Theses, and Masters Projects

No abstract provided.


Studies On The Role Of Expression Of Phosphoenolpyruvate Carboxykinase Gene In The Development Of Bovine Lactation Ketosis , Sowmya Moorthi Jan 1992

Studies On The Role Of Expression Of Phosphoenolpyruvate Carboxykinase Gene In The Development Of Bovine Lactation Ketosis , Sowmya Moorthi

Retrospective Theses and Dissertations

Changes in expression of the phosphoenolpyruvate carboxykinase (PEPCK) gene, which codes for a key regulatory enzyme of gluconeogenesis, were studied in bovine liver tissue. Induction of ketosis was attempted in cows by a protocol of restricting feed intake (FR) and supplementing with 1,3-butanediol (BD), a ketogenic precursor, beginning at 15 d postpartum (FRBD protocol). Twenty Holstein cows were distributed in equal numbers among four treatments: normal control, obese control, normal cows given FRBD, and obese cows given FRBD. Liver biopsies were obtained at nine different times during early lactation: 10 d prepartum, 5 and 14 d postpartum (just before ...


Cloning And Characterization Of Mink Plasminogen Activator Inhibitor Type 1 (Pai-1) Cdna And The Regulation Of Mink Pai-1 Expression At Mrna Level , Tsung-Hsien Chuang Jan 1992

Cloning And Characterization Of Mink Plasminogen Activator Inhibitor Type 1 (Pai-1) Cdna And The Regulation Of Mink Pai-1 Expression At Mrna Level , Tsung-Hsien Chuang

Retrospective Theses and Dissertations

Mink plasminogen activator inhibitor type 1 (PAI-1) is a glycoprotein with molecular weight about 50 kD and is highly regulated in cultured cells. In mink lung CCL64 epithelial cells, epidermal growth factor (EGF), transforming growth factor-[beta] (TGF-[beta]), and 12-O-tetradecanoyl phorbol-13-acetate (TPA) increase the synthesis of PAI-1. Epidermal growth factor and TGF-[beta] synergistically stimulate the synthesis of PAI-1. The synergistic effect and factors which govern the PAI-1 expression in mink lung CCL64 cells were studied by using a mink PAI-1 cDNA as a probe to detect PAI-1 mRNA;The mink PAI-1 cDNA was cloned from a cDNA library ...


Parent Selection To Maximize Heterosis Expression In Soybean (Glycine Max L. [Merr.]) , Francisco Jonathan Cerna Jan 1992

Parent Selection To Maximize Heterosis Expression In Soybean (Glycine Max L. [Merr.]) , Francisco Jonathan Cerna

Retrospective Theses and Dissertations

Utilization of heterosis effects in breeding soybean (Glycine max (L.) Merr) cultivars for commercial production is still a possibility. One objective of this research was to determine heterosis expression in soybean germplasms obtained from the crossing of parents selected on the basis of restriction fragment length polymorphism (RFLP) diversity and isozyme loci (ISO) constitution compared to parents selected for high-yielding (HY) ability and differences in geographic origin (GO). The second objective was to compare mean heterotic performance of F[subscript]1 hybrids and F[subscript]2 bulks evaluated in hill- and row-plots;Twenty-one genotypes of each of Maturity Groups (MG ...


Translational Frameshifting By Barley Yellow Dwarf Virus Rna , Rong Di Jan 1992

Translational Frameshifting By Barley Yellow Dwarf Virus Rna , Rong Di

Retrospective Theses and Dissertations

Barley yellow dwarf virus is a single-stranded RNA virus, consisting of several serotypes including the PAV serotype which is the focus of this dissertation. The 39K open reading frame (ORF) at the 5[superscript]' end of the 5.7 kb (kilobase) genome overlaps with a downstream 60K ORF by 13 nucleotides. Several approaches were employed to demonstrate that the 60K ORF (putative polymerase gene) is expressed by an unusual minus one (-1) translational frameshifting mechanism. A sequence encompassing this overlap region induces frameshifting in heterologous and native contexts. The efficiency of frameshifting induced by the PAV sequence was approximately 3 ...


Molecular Analysis Of Two Adjacent Transcription Terminators In Yeast , Julie A. Peterson Jan 1992

Molecular Analysis Of Two Adjacent Transcription Terminators In Yeast , Julie A. Peterson

Retrospective Theses and Dissertations

RHO1, a member of the ras superfamily of genes, is located on chromosome XIII adjacent to MRP2, a gene coding for mitochondrial ribosomal protein (Madaule and Axel, 1985, Myers, et al. 1987) in Saccharomyces cerevisiae. Nucleotide sequence analysis demonstrated that RHO1 and MRP2 are transcribed convergently and the 3[superscript]' ends of the two coding sequences are separated by only 281 nucleotides (Madaule et al., 1987);The physical relationship between RHO1 and MRP2 was investigated at the transcriptional level by mapping the termini of both transcripts. RHO1 transcripts were found to possess extensive nontranslated regions with jagged initiation and polyadenylation ...


Direct Introduction Of Dna Into Embryonic Axes Of Glycine Max By Microprojectile Bombardment , Charisse Marie Buising Jan 1992

Direct Introduction Of Dna Into Embryonic Axes Of Glycine Max By Microprojectile Bombardment , Charisse Marie Buising

Retrospective Theses and Dissertations

Apical meristem and axillary bud development in germinating soybean embryonic axes treated for 24 hr with the cytokinin, 6-benzylaminopurine (BAP) seemed normal morphologically, but was delayed by about 9 to 15 hr. After 5 days, numerous, rapidly proliferating meristematic regions (de novo vegetative buds) were observed in BAP-treated axes around the perimeter of the apical dome. These visible changes were preceded by transient inhibition of DNA synthesis in the shoot apices as early as 21-36 hr post-imbibition;Some of the new meristematic regions developed into multiple shoots, which were excised, subcultured, and used to generate mature plants. In the absence ...


Fluorescence Detection For Gel And Capillary Electrophoresis , Barry L. Hogan Jan 1992

Fluorescence Detection For Gel And Capillary Electrophoresis , Barry L. Hogan

Retrospective Theses and Dissertations

Several separation and detection schemes for gel and capillary electrophoresis are described. Emphasis is placed upon direct and indirect fluorescence detection systems as utilized for the determination of proteins, peptides, and intracellular analytes;First, an indirect fluorescence detection system for the separation of proteins via gel electrophoresis is described. Quantities as low as 50 nanograms of bovine serum albumin and soybean trypsin inhibitor are separated and detected visually without the need for staining of the analytes. This is very similar to levels of protein commonly separated with gel electrophoresis;A separation and detection system utilizing indirect fluorescence detection with capillary ...


Novel Detection Schemes And Automated Image Analysis Algorithms For Planar Chromatography And Gel Electrophoresis , Lance Bryant Koutny Jan 1992

Novel Detection Schemes And Automated Image Analysis Algorithms For Planar Chromatography And Gel Electrophoresis , Lance Bryant Koutny

Retrospective Theses and Dissertations

After a detailed introduction to Charge Coupled Devices (CCDs) and a discussion of personal computer capabilities, examples of their applications involving novel analytical techniques are presented;A new laser-based indirect fluorometric detection method for thin-layer chromatography is demonstrated with digitally controlled two-dimensional scanning and computer-aided image acquisition and vision. Utilizing this mode, universal detection is possible without derivatization, with a detection limit of 6 pg; 100 times lower than with human vision;A detection scheme for DNA in agarose slab gel electrophoresis based on ultraviolet absorption allows for convenient in situ monitoring of conventional gels via a CCD camera system ...


Association Of Tetrapyrrole Intermediates In The Bacteriochlorophyll A Biosynthetic Pathway With The Major Outer-Membrane Porin Protein Of Rhodobacter Capsulatus, David W. Bollivar, Carl E. Bauer Dec 1991

Association Of Tetrapyrrole Intermediates In The Bacteriochlorophyll A Biosynthetic Pathway With The Major Outer-Membrane Porin Protein Of Rhodobacter Capsulatus, David W. Bollivar, Carl E. Bauer

David Bollivar

Rhodobacter capsulatus regulates synthesis of bacteriochlorophyll a in response to changes in oxygen partial pressure and light intensity. One early model proposed that this regulation involved a carrier polypeptide that functions to tether tetrapyrrole intermediates to the membrane. In the present study we isolated tetrapyrrole intermediates accumulated in three strains of R. capsulatus that contain mutations which block bacteriochlorophyll a biosynthesis at different steps of the magnesium branch of the pathway. Each of the tetrapyrrole intermediates was shown to be associated with the same 32 kDa polypeptide, as indicated by similar electrophoretic mobility and antigenic cross-reactivity with polyclonal antisera. The ...


Glossary For Chemists Of Terms Used In Biotechnology, B. Nagel, H. Dellweg, Lila Gierasch Dec 1991

Glossary For Chemists Of Terms Used In Biotechnology, B. Nagel, H. Dellweg, Lila Gierasch

Lila Gierasch

The glossary contains definitions and explanatory notes, if needed, of over 230 terms frequently used in publications in the multidisciplinary field of biotechnology. The glossary was developed as a step to help facilitate communication between chemists, chemical engineers, biologists and bioengineers and to make biotechnology and its methodologies more accessible to the chemical profession. The interrelation between chemistry, chemical engineering and the burgeoning areas of molecular biology will be especially important as chemical industrial processes begin to incorporate recombinant DNA techniques, for example. The range of terms includes microbiology, genetic engineering, biochemistry, molecular biology, biochemical engineering, bioprocessing and general concepts ...


Nucleotide Sequence Of S-Adenosyl-L-Methionine: Magnesium Protoporphyrin Methyltransferase From Rhodobacter Capsulatus, David W. Bollivar, Carl E. Bauer Dec 1991

Nucleotide Sequence Of S-Adenosyl-L-Methionine: Magnesium Protoporphyrin Methyltransferase From Rhodobacter Capsulatus, David W. Bollivar, Carl E. Bauer

David Bollivar

No abstract provided.