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Biochemical Analysis Of Putative Rna Binding Proteins In Porphyromonas Gingivalis
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Book Synopsis Biochemical Analysis of Putative RNA-binding Proteins in Porphyromonas Gingivalis by : Jacob Mirpanah
Download or read book Biochemical Analysis of Putative RNA-binding Proteins in Porphyromonas Gingivalis written by Jacob Mirpanah and published by . This book was released on 2018 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: RNA-Binding Proteins (RBPs) play important regulatory roles in countless cellular processes. Often via the induction of a structural change in RNA's secondary structure, RBPs are known to modulate protein expression through post-transcriptional regulation. In Escherichia coli, such RBPs have been thoroughly studied and shown to display differential expression throughout the bacterial life cycle; suggesting their importance in prompting different events in the bacterial cell. Further, RNA- and DNA-recognition domains have been characterized in proteins Hub as well as RBP in E.coli and their nucleic acid ligands sequenced. It was our aim to extend this level of information to Hub and RBP as they exist in a main etiological agent in periodontal disease; Porphyromonas gingivalis. Using quantitative PCR expression analysis, we saw a general upregulation of both proteins in the logarithmic phase as compared to the stationary phase. This upregulation was most pronounced in PG0627, the putative analog of E. coli RBP in P. gingivalis. Further, Electrophoretic Mobility Shift Assays suggest sequence specific interaction of PG0627 to RNA. Uninhibited and Inhibited mobility assays seem to confirm that PG0627 binds with great specificity to a conserved RNA sequence. Quantitative measure of interaction came in the form of fluorescence anisotropy, which produced a Dissociation Constant (Kd) of approximately 53 nM; suggesting a high degree of affinity. Lastly, a mutant was generated in order to produce high quality RNA libraries to be sequenced through the Illumina MiSeq system. Sequencing data is still incoming.
Book Synopsis Biochemical Analysis of Putative Single-stranded Nucleic Acid Binding Proteins in Porphyromonas Gingivalis by : Steve H. Kokorelis
Download or read book Biochemical Analysis of Putative Single-stranded Nucleic Acid Binding Proteins in Porphyromonas Gingivalis written by Steve H. Kokorelis and published by . This book was released on 2017 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Proteins that bind to both DNA and RNA embody the ability to perform multiple functions by a single gene product. These nucleic acid binding proteins in prokaryotes can play a vital role in many cellular processes, including replication, transcription, gene expression, recombination, and repair, to name a few. Nucleic acid binding proteins have unique functional characteristics that stem from their structural attributes that have evolved in a widely-conserved manner. In Escherichia coli (E. coli), the highly-conserved histone-like protein, HU, which predominates as a heterodimer of HU[alpha] and HU[beta], has been found to bind to both dsDNA and ssDNA. Likewise, RNA-binding proteins contain various structural motifs, many of which are also conserved amongst many bacterial species like the RNA recognition motif. However, in Porphyromonas gingivalis, a periodontal pathogen, the histone-like, HU proteins and the RNA-binding protein (RBP) are not well characterized compared to their respective structures in E. coli. In our study, we sought to characterize and compare the HU proteins and RBP in order to gain a better understanding of their structure and function in the cell. Our data showed the HU proteins predominate as homo-tetramers and RBP as a monomer. We demonstrated single-stranded DNA binding with all three proteins. We found both P. gingivalis HU subunits bind non-specifically to ssDNA but show preferential binding to poly(dG) content, while binding to poly(dA) the weakest. These results show that HU[alpha], HU[beta] and RBP are novel ssDNA binding proteins in P. gingivalis, indicating an expanded role and function within the cell.
Book Synopsis Characterization of the Biological Role of a Putative Porphyromonas Gingivalis RNA-binding Protein by : Romana Cvitkovic
Download or read book Characterization of the Biological Role of a Putative Porphyromonas Gingivalis RNA-binding Protein written by Romana Cvitkovic and published by . This book was released on 2014 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Porphyromonas gingivalis, a gram-negative anaerobic bacterium, is a major etiological agent in the initiation and progression of severe forms of periodontal disease. Oral bacteria like P. gingivalis are subject to continually changing conditions as a consequence of host eating, oral hygiene patterns and subgingival temperatures. As such survival requires an adaptive response to environmental cues, but little is known about the mechanism by which P. gingivalis controls co- and post-transcriptional regulation of RNA levels and potentially protein expression. RNA-binding proteins (RBPs) are evolutionarily conserved across species and are involved in such regulatory mechanisms. However, P. gingivalis currently has no identified RBP. Recently, PG0627 has become an ideal candidate for a putative RBP due to its sequence homology to RBPs across various species. By characterizing PG0627, we can gain better insight into the function of this hypothetical protein and determine if it indeed behaves like an RNA-binding protein. A host of studies were done on a PG0627-deficient P. gingivalis mutant, V3139, in order to determine the biological role of the protein encoded by the gene. Our bioinformatics analysis indicated that PG0627 had sequence homology to several RNA recognition motifs or RBPs. Furthermore, our PG0627-deficient mutant, when compared to W83, exhibited decreased cell-associated iron content, decreased total interactions and invasions with eukaryotic cells, and decreased protease activity. Conversely, our PG0627-deficient mutant displayed slightly increased growth in the presence of nitrosative stress, and in hemin-depleted conditions. In conclusion, our results support that PG0627 is a valid candidate for an RNA-binding protein in P. gingivalis.
Book Synopsis Characterization of Putative Porphyromonas Gingivalis RNA-binding Proteins by : Holly Dwyer
Download or read book Characterization of Putative Porphyromonas Gingivalis RNA-binding Proteins written by Holly Dwyer and published by . This book was released on 2014 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: Porphyromonas gingivalis (P. gingivalis) is a gram-negative, anaerobic bacterium recognized as a major player in progression of periodontal disease. P. gingivalis survives in the oral cavity while being exposed to dynamic environmental conditions such as pH, temperature, nutrient availability and host immune responses such as oxygen tension and nitrosative stress. Survival and pathogenesis of P. gingivalis in the oral cavity require mechanisms to regulate gene expression in response to the extracellular signals. Little is known about the regulatory mechanisms of P. gingivalis in the oral cavity, so it is important to investigate and characterize these regulatory mechanisms. Adaptation to environmental cues using riboregulation is a significant mechanism for post-transcriptional regulation in bacteria. Using bioinformatics, we have identified a putative RNA-binding protein in P. gingivalis: RBP. Bioinformatic studies have led to the selection of HU[beta] and HU[alpha]nucleoid associated proteins as controls for RNA binding. I hypothesize that the candidate proteins RBP, HU[beta] and HU[alpha]bind RNA in P. gingivalis. The first aim is to show that RBP, HU[beta] and HU[alpha]bind RNA. Using electrophoretic mobility shift assays with IRE RNA and synthesized RNA motifs, I have confirmed that the proteins do bind RNA. The second aim is to isolate and sequence the P. gingivalis RNA that bind to RBP, HU[beta] and HU[alpha]. I have isolated the RNAs that bound the proteins and determined identity of the RNA using high throughput sequencing. Finally, I have identified an antibody that specifically binds RBP to use for in vivo immunoprecipitation of RNA-protein complexes from P. gingivalis. In conclusion RBP, HU[beta] and HU[alpha]are novel RNA binding proteins in P. gingivalis, and further investigation of these proteins is necessary to understand the mechanisms of gene regulation in P. gingivalis.
Book Synopsis Polymicrobial Diseases by : Kim A. Brogden
Download or read book Polymicrobial Diseases written by Kim A. Brogden and published by . This book was released on 2002 with total page 460 pages. Available in PDF, EPUB and Kindle. Book excerpt: Polymicrobial diseases, those involving more than one etiologic agent, are more common than is generally realized and include respiratory diseases, gastroenteritis, conjunctivitis, keratitis, hepatitis, periodontal diseases, multiple sclerosis, genital infections, intra -- abdominal infections, and pertussis.
Book Synopsis Cambridge Scientific Biochemistry Abstracts by :
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Book Synopsis Current Advances in Protein Biochemistry by :
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Book Synopsis Bacterial Pathogenesis by : Brenda A. Wilson
Download or read book Bacterial Pathogenesis written by Brenda A. Wilson and published by John Wiley & Sons. This book was released on 2020-08-11 with total page 683 pages. Available in PDF, EPUB and Kindle. Book excerpt: This highly anticipated update of the acclaimed textbook draws on the latest research to give students the knowledge and tools to explore the mechanisms by which bacterial pathogens cause infections in humans and animals. Written in an approachable and engaging style, the book uses illustrative examples and thought-provoking exercises to inspire students with the potential excitement and fun of scientific discovery. Completely revised and updated, and for the first time in stunning full-color, Bacterial Pathogenesis: A Molecular Approach, Fourth Edition, builds on the core principles and foundations of its predecessors while expanding into new concepts, key findings, and cutting-edge research, including new developments in the areas of the microbiome and CRISPR as well as the growing challenges of antimicrobial resistance. All-new detailed illustrations help students clearly understand important concepts and mechanisms of the complex interplay between bacterial pathogens and their hosts. Study questions at the end of each chapter challenge students to delve more deeply into the topics covered, and hone their skills in reading, interpreting, and analyzing data, as well as devising their own experiments. A detailed glossary defines and expands on key terms highlighted throughout the book. Written for advanced undergraduate, graduate, and professional students in microbiology, bacteriology, and pathogenesis, this text is a must-have for anyone looking for a greater understanding of virulence mechanisms across the breadth of bacterial pathogens.
Book Synopsis Index to Theses with Abstracts Accepted for Higher Degrees by the Universities of Great Britain and Ireland and the Council for National Academic Awards by :
Download or read book Index to Theses with Abstracts Accepted for Higher Degrees by the Universities of Great Britain and Ireland and the Council for National Academic Awards written by and published by . This book was released on 2008 with total page 308 pages. Available in PDF, EPUB and Kindle. Book excerpt:
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Author :Paul E. Kolenbrander Publisher :American Society for Microbiology Press ISBN 13 :1555815030 Total Pages :617 pages Book Rating :4.5/5 (558 download)
Book Synopsis Oral Microbial Communities by : Paul E. Kolenbrander
Download or read book Oral Microbial Communities written by Paul E. Kolenbrander and published by American Society for Microbiology Press. This book was released on 2011-08-02 with total page 617 pages. Available in PDF, EPUB and Kindle. Book excerpt: Understand how the intricacies of multispecies community life are related to human oral health. * Explores the immense opportunities presented by readily accessible, genetically tractable, genome-sequenced oral species that naturally form multispecies communities. * Highlights model systems that study oral bacterial interactions, including biofilm growth using saliva as the source of nutrition. * Emphasizes the use of genomic inquiry to probe the human oral microbiome.
Book Synopsis Prokaryotic Cell Wall Compounds by : Helmut König
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