244 research outputs found
Announcement of the 2019 BLAST Conference: “BLAST XV: 15th International Conference on Bacterial Locomotion and Signal Transduction”
No full textABSTRACT An exciting conference showcasing cutting edge research in bacterial signal transduction, chemotaxis, and motility will be held in January 2019. This conference, called Bacterial Locomotion and Signal Transduction (BLAST), will be held in New Orleans, LA, USA, under the auspices of chair Birgit Scharf. The conference has been held biennially since 1991 and highlights presentations from early-career scientists. The majority of talks are from submitted abstracts, interspersed by presentations of internationally renowned scientists. Six awards will be presented specifically to young researchers. The main goal is to bring together researchers at the junior and experienced levels from a wide variety of research disciplines. Topics showcased include structural biology, computational biology, modeling, and cell biology covering molecular mechanisms of bacterial movement, systems biology, evolution of signaling systems, computational modeling of nanomachine function, and bacterial-host interactions. The combination of broad-scope and in-depth science will continue to inspire breakthrough technical and scientific advances
Bibliographics for the 983 eprints in the live archives of E-LIS : trends and status report up to 7th July 2004, based on author-self-archiving metadata
Get PDFThe priority for ideas and philosophy related to "Network Theory" have been traced back and documented by Braun(2004),and credit goes to Karinthy(1929).The IT has empowered to realise it, as the most practical phenomena and it is no more a humour. The OAI (Open Archives Initiatives)and ACIS (Academic Contributor Information System)are progressive in the direction ,which may lead to realise the "Collective Genius" at global level. Focus of present study is on Author-Self-Archiving (A-S-A)Metadata of the 983 Eprints in the Live Archives of the E-LIS (EPrints of Library and Information Science),which were approved till 7th July 2004.The A-S-A Metadata was used for librametric analysis. Self-explanatory bibliographics are illustrated.The highlights include: Conference papers (34%); highest approval, June 2004 (28%); published archives (76%);not refereed (52%); not in public domain (60%); highest self-archiving-author (De Robbio, Antonella).The Nos. of EPrints having single JITA domain specifications were: Theoretical and general aspects of libraries and information(27); Information use and sociology of information(80);Users,literacy and reading(13);Libraries as physical collections(30);Publishing and legal issues(57);Management(13);Industry, profession and education(36);Information sources, supports, channels(113) ; Information treatment for information services, Information functions and techniques (101); Technical services libraries, archives and museums(25); Housing technologies(1); Information technology and library technology(92); and Inter-domainery (395) i.e. having specifications of two or more than two JITA classes
Comparative Analysis of Flagellotropic Agrobacterium Phages
No full textThe planetary microbiome is dominated by bacteriophages, the viruses of bacteria. Bacteriophages utilize various means to infect their bacterial hosts, but all phages require receptors on the host surface. Flagellotropic bacteriophages involve an actively rotating bacterial flagellum to attach and initiate the infection process. Flagellotropic bacteriophages are poorly studied compared to their non-flagellotropic counterparts. It is important to research the relationship between these phages and their hosts to elucidate the mechanisms behind the attachment and infection process. It is imperative that we expand the knowledge on these mechanisms due to the increase in potential phage applications both medically and environmentally. This study investigates three flagellotropic phages and their bacterial hosts: Milano and host Agrobacterium tumefaciens str. C58, OLIVR4 and host Agrobacterium Biovar 1 str. ST15.13/040, and 7-7-1 and host Agrobacterium sp. H13-3.
Chapter 1 is a literature review. Chapter 2 discusses the identification of the bacterial lipopolysaccharide as a secondary receptor for bacteriophage Milano, and the cross infectivity of the three bacteriophages with the three host bacteria. We quantified infectivity of the three bacterial host strains with the three bacteriophages by performing Efficiency of Plating assays (EOPs) and observed interesting relationships between the phages and host strains. We were also able to further support the hypothesis that OLIVR4, a recently discovered Agrobacterium phage, is indeed flagellotropic by infecting a host mutant with non-rotating flagella with OLIVR4. Overall, this research will help expanding the knowledge of this special class of bacteriophages, opening the door for their further applications in medicine and environmental sciences.Master of ScienceBacteriophages, phages for short, are viruses that only infect bacteria. While phages can use many methods to infect their hosts, all of these methods require receptors- a structure that is produced by the bacterial host cell – for attachment. Phages that use the host flagellum, which is a helical filament that protrudes from the bacterial cell and facilitates movement, are referred to as flagellotropic phages. For these phages, an actively rotating bacterial flagellum is a requirement for infection, but there can be additional structures that aide in adhesion and infection. These additional structures are referred to as secondary receptors. Phages can be used to supplement or even replace antibiotic treatment, both medically and environmentally. As antibiotic resistance is a rising concern in the medical sector especially, phages are rising as a treatment option. One benefit of the use of phages is their high specificity, which enables them to kill only the target bacterium while leaving the rest of the microbiome intact.
In this work, we investigated three different flagellotropic phages—Milano, OLIVR4, and 7-7-1. First, we identified secondary receptors for Milano. We also studied how the three phages behaved when they were given the opportunity to infect different Agrobacterium species. Agrobacteria are known to cause Crown gall disease—a disease that results in tumor-like growths in plants. It is important to understand how these phages interact with their hosts to work towards potential therapeutics for this devastating plant disease
Characterization of the thioredoxin system in Methanosarcina mazei
No full textThioredoxin (Trx) and thioredoxin reductase (TrxR) along with an electron donor form a thioredoxin system. Such systems are widely distributed among the organisms belonging to the three domains of life. It is one of the major disulfide reducing systems, which provides electrons to several enzymes, such as ribonucleotide reductase, methionine sulfoxide reductase and glutathione peroxidase to name a few. It also plays an important role in combating oxidative stress and redox regulation of metabolism. Trx is a small redox protein, about 12 kDa in size, with an active site motif of Cys-X-X-Cys. The reduction of the disulfide in Trx is catalyzed by TrxR. Two types of thioredoxin reductases are known, namely NADPH thioredoxin reductase (NTR) with NADPH as the electron donor and ferredoxin thioredxoin reductase (FTR) which depends on reduced ferredoxin as electron donor. Although NTR is widely distributed in the three domains of life, it is absent in some archaea, whereas FTRs are mostly found in plants, photosynthetic eukaryotes, cyanobacteria, and some archaea.
The thioredoxin system has been well studied in plants, mammals, and a few bacteria, but not much is known about the archaeal thioredoxin system. Our laboratory has been studying the thioredoxin systems of methanogenic archaea, and a major focus has been on Methanocaldococcus jannaschii, a deeply rooted archaeon that has two Trxs and one TrxR. My thesis research concerns the thioredoxin system of the late evolving members of the group which are exposed to oxygen more frequently than the deeply rooted members of the group, and have several Trxs and TrxRs. Methanosarcina mazei is one such organism, whose thioredoxin system is composed of one NTR, two FTRs, and five Trx homologs.
Characterization of the components of a thioredoxin system sets the basis to further explore its function. I have expressed in Escherichia coli and purified the five Trxs and three TrxRs of M. mazei. I have shown the disulfide reductase activities in MM_Trx1 and MM_Trx5 by their ability to reduce insulin with DTT as the electron donor, and that in MM_Trx3 through the reduction of DTNB by this protein with NADPH as the electron donor, and in the presence of NTR as the enzyme. MM_Trx3 was found to be the only M. mazei thioredoxin to accept electrons through the NTR, and to form a complete Trx - NTR system. The Trx - FTR systems are well studied in plants, and such a system is yet to be defined in archaea. I have proposed a mechanism of action for one of the FTRs. FTR2 harbors a rubredoxin domain, and this unit is the only rubredoxin in this organism. Superoxide reductase, an enzyme that reduces superoxide radical to hydrogen peroxide without forming oxygen, utilizes rubredoxin as the direct electron source and this enzyme is found in certain anaerobes, including Methanosarcina species. Thus, it is possible that FTR2 provides electrons via a Trx to the superoxide reductase of M. mazei. This activity will define FTR2 as a tool in combating oxidative stress in M. mazei.
In my thesis research I have laid a foundation to understand a complex thioredoxin system of M. mazei, to find the role of each Trx and TrxR, and to explore their involvement in oxidative stress and redox regulation.Master of Scienc
Molecular Microbiology
Get PDFSensory adaptation in bacterial chemotaxis is mediated by posttranslational modifications of methyl-accepting chemotaxis proteins (MCPs). In Escherichia coli, the adaptation proteins CheR and CheB tether to a conserved C-terminal receptor pentapeptide. Here,we investigated the function of the pentapeptide motif (N/D)WE(E/N)F in Sinorhizobium meliloti chemotaxis. Isothermal titration calorimetry revealed stronger affinity of the pentapeptides to CheR and activated CheB relative to unmodified CheB. Strains with mutations of the conserved tryptophan in one or all four MCP pentapeptides resulted in a significant decrease or loss of chemotaxis to glycine betaine, lysine, and acetate, chemoattractants sensed by pentapeptide-bearing McpX and pentapeptide-lacking McpU and McpV, respectively. Importantly, we discovered that the pentapeptide mediates chemotaxis when fused to the C-terminus of pentapeptide-lacking chemoreceptors via a flexible linker. We propose that adaptational assistance and a threshold number of available sites enable the efficient docking of adaptation proteins to the chemosensory array. Altogether, these results demonstrate that S. meliloti effectively utilizes a pentapeptide-dependent adaptation system with a minimal number of tethering units to assist pentapeptide-lacking chemoreceptors and hypothesize that the higher abundance of CheR and CheB in S. meliloti compared to E. coli allows for ample recruitment of adaptation proteins to the chemosensory array.Published versio
Similarities and variations of the enterobacterial chemotaxis paradigm in Sinorhizobium meliloti
No full textSinorhizobium meliloti is a nitrogen-fixing endosymbiont of the legume Medicago sativa commonly known as alfalfa. It uses flagellar rotation and chemotaxis to seek roots of host plants to inhabit. This symbiosis serves as a great model system for studying biological nitrogen fixation and plant-microbe interactions. Since alfalfa brings enormous economic value to the USA, investments into the knowledge of the chemotaxis process that initiates symbiosis have the ability to mitigate deterioration of the environment and significantly increase food supply. The chemotaxis system in the enteric bacteria Escherichia coli is well studied and has been a great resource to understanding the process in other bacterial systems including our model organism S. meliloti.
This dissertation compares and contrasts the chemotaxis features in E. coli and S. meliloti and investigates their molecular functions. Based on the understanding gained so far, we attempt to offer plausible explanations for the underlying mechanisms of the S. meliloti chemotaxis pathway. Chapter 1 describes why biological nitrogen fixation is important for agriculture and the health of our environment. This chapter also sheds light on the symbiotic relationship between alfalfa and S. meliloti, which culminates in the formation of nitrogen fixing nodules. We expound on the chemotaxis systems in E. coli and other bacteria including S. meliloti and Bacillus subtilis.
In chapter 2, we compare the distribution of C-terminal pentapeptide-bearing receptors and the adaptation proteins that they tether in E. coli and S. meliloti. The stoichiometry data show that the ratio of pentapeptide-bearing chemoreceptors to chemotaxis protein (Che)R and CheB molecules are approximately 500- and 160-fold higher in S. meliloti than in E. coli, respectively. Since not all chemoreceptors in chemotactic bacteria have and utilize the pentapeptide moiety, we investigated the S. meliloti system and observed a strong interaction between CheR, activated CheB and the isolated pentapeptides via in-vitro binding studies. On the contrary, unmodified CheB showed weak binding to the pentapeptide. Through in-vivo studies, we highlighted the physiological necessity of the pentapeptide for chemotaxis. S. meliloti strains with substitutions of the conserved tryptophan residue to alanine in one or all four pentapeptide-bearing Methyl-accepting Chemotaxis Proteins (MCPs) resulted in diminished or loss of chemotaxis to glycine betaine, lysine, and acetate, ligands sensed by pentapeptide-bearing McpX and pentapeptide-lacking McpU and McpV, respectively. The flexible linker connecting the pentapeptide to the MCPs together with the pentapeptide itself were shown to be functional on pentapeptide-lacking chemoreceptors and provided adaptational assistance to other chemoreceptors that lacked a functional pentapeptide. Based on these results, we concluded that S. meliloti employs a pentapeptide-dependent adaptation system with MCPs possessing a consensus pentapeptide motif (N/D)WE(E/N)F). Finally, we postulated that the higher abundance of CheR and CheB in S. meliloti compared to E. coli compensates for the lower number of pentapeptide-bearing chemoreceptors in the chemosensory array.
In chapter 3, we explored the putative phosphatase function of a novel protein, CheT, on phosphorylated S. meliloti response regulators. The kinase CheA phosphorylates both the sink response regulator, CheY1, and the flagellar motor interacting response regulator, CheY2. CheY1 competes with CheY2 for these phosphate groups, but we have discovered another layer of complexity to the story. Sequence comparison of S. meliloti CheT and the E. coli phosphatase CheZ shows little sequence homology. However, both proteins share a DXXXQ phosphatase motif. Phosphorylation assays performed using radiolabeled [γ-32P]-ATP revealed that CheT acts as a phosphatase of CheY1~P and accelerates dephosphorylation of CheY1~P by at least two-fold. Interestingly, we also discovered that CheT interacts with CheR, but this interaction did not affect the enzymatic activity of either protein under the examined conditions. Unexpectedly, a cheT deletion strain and strains carrying mutations in the phosphatase motif exhibit an increased swimming speed, a phenotype that does not conform with the model that the absence of CheT or its activity results in increased CheY2~P levels and reduced swimming speed. We concluded that a revised S. meliloti signal termination pathway should include CheT enhancing dephosphorylation of CheY1~P and sensory adaptation involving the yet unknown function of CheT on CheR.
While the adaptation system in S. meliloti is unexplored, this work provides first insights into fascinating deviations and similarities to the known paradigm. We have also delivered evidence that the S. meliloti signal termination system requires a dedicated phosphatase. The knowledge gained here takes us a step closer to enhance the S. meliloti chemotaxis pathway towards improved symbiosis with alfalfa and to reduce our dependence on environmentally deleterious synthetic fertilizers.Doctor of PhilosophyLike all living things, bacteria inhabit a constantly changing environment, hence the need to take up and process this information. Bacterial cells have evolved sophisticated biological tools to tackle this challenge of detecting, responding and adapting to environmental signals like nutrients, toxins, temperature changes, light, metabolites, etc. Motile bacteria such as Escherichia coli, a gut resident microbe, and Sinorhizobium meliloti, a soil dwelling bacterium, direct their swimming behavior in response to chemical gradients within the milieu through a process termed chemotaxis. Generally, this vital process enables a bacterium to escape harmful chemicals and gravitate towards beneficial ones. However, S. meliloti specifically employs chemotaxis to locate the roots of its plant host (alfalfa) and to establish a symbiotic relationship through which the bacteria provide essential nitrogen for plant growth in exchange for nourishment. The biological tools employed by S. meliloti for chemotaxis include environmental sensing receptors called Methyl-accepting Chemotaxis Proteins (MCPs) and proteins inside the bacterial cell that transfer information from the sensors to long, helical rotating propeller structures, called flagella. Importantly, the efficiency of this process hinges on a timely termination of information flow and the ability to adapt to prevailing stimuli while maintaining sensitivity to increasing concentration gradients. This work investigates the function of the C-terminal five amino acid motif of MCPs known to be critical for adaptation in E. coli and the phosphatase activity of a novel protein, CheT, in signal termination of S. meliloti chemotaxis system
La "máquina de trovar" de Antonio Machado o cómo la técnica sustituye al poeta
Get PDFAntonio Machado, the singer of the dry lands of Castille, discovers an unexpected side of the magic secrets of the modern tecnic in his text about; the "machine of versifying" from his "Cancionero apócrifo" (1926/28). The analysis of this text leads to the centre of an interest which is particularly modern; the worrying power of the media and the sustitution of the author/subject for a machine. This essay looks into the prehistory of the interactive media and at the same time allows a new glance at Antonio Machado's position in the "98 generation".Antonio Machado, le chantres des terres arides de la Castille, découvre un penchant inattendu pour les Secrets magiques de la technique moderne dans son texte sur la "machine â trouver" de son "Cancionero apócrifo" (1926/28). L'analyse de ce texte porte au centre d'un trouble proprement moderne: le pouvoir inquiétant des médias et la substitution de 1' auteur/sujet par une machine. L'essai de Birgit Wagner exploit la préhistoire des médias interactifs et permet en même temps un regard nouveau sur la position d'Antonio Machado au sein de la "génération de 98".Publicad
Identification of Receptor Binding Proteins in Flagellotropic <i>Agrobacterium</i> Phage 7-7-1
No full textThe rapid discovery of new and diverse bacteriophages has driven the innovation of approaches aimed at detailing interactions with their bacterial hosts. Previous studies on receptor binding proteins (RBPs) mainly relied on their identification in silico and are based on similarities to well-characterized systems. Thus, novel phage RBPs unlike those currently annotated in genomic and proteomic databases remain largely undiscovered. In this study, we employed a screen to identify RBPs in flagellotropic Agrobacterium phage 7-7-1. Flagellotropic phages utilize bacterial flagella as receptors. The screen identified three candidate RBPs, Gp4, Gp102, and Gp44. Homology modelling predicted that Gp4 is a trimeric, tail associated protein with a central β-barrel, while the structure and function of Gp102 and Gp44 are less obvious. Studies with purified Gp41-247 confirmed its ability to bind and interact with host cells, highlighting the robustness of the RBP screen. We also discovered that Gp41-247 inhibits the growth of host cells in a motility and lipopolysaccharide (LPS) dependent fashion. Hence, our results suggest interactions between Gp41-247, rotating flagellar filaments and host glycans to inhibit host cell growth, which presents an impactful and intriguing focus for future studies
Characterization of the Sinorhizobum Meliloti Chotaxis System
Get PDFIncreasing awareness to global climate change has drastically focused attention on finding solutions to reduce environmental impacts while still providing sufficient food for the increasing world population. Beneficial Nitrogen Fixing (BNF) microbes provide a possible solution by delivering biological nitrogen to plants resulting in reduced environmental impacts due to fertilizer runoff and eutrophication. One well studied model is that of Sinorhizobium meliloti and its legume host Medicago sativa (alfalfa), the fourth largest USA crop used for animal feed. Advancing research for this symbiosis model can provide solutions to enhance yield while minimizing environmental impacts.
Chapter 2 focuses on the deviation of the S. meliloti chemotaxis system from the enteric paradigm. Quantitative immunoblots determined the cellular amounts of chemotaxis proteins. Overall, chemotaxis protein levels were approximately 10-fold lower in S. meliloti compared to B. subtilis and E. coli. Focusing on cellular stoichiometric ratios, S. meliloti generally exhibits drastically higher values for CheB, CheR, and CheY to the histidine kinase CheA monomer compared to E. coli and B. subtilis.
Chapter 3 characterizes the role of McpX to quaternary ammonium compound (QAC) sensing. QACs are exuded by germinating alfalfa seeds. In vitro binding assays were performed to determine ligand binding characteristics. S. meliloti chemotaxis behavior to QACs was analyzed in in vivo capillary assays under real-time imaging.
These studies strengthen our knowledge of the chemotaxis system in the symbiosis model of S. meliloti and alfalfa. The data can further be used to create a mathematical model of the dynamics of bacteria-host interaction. The results can be used to optimize chemotaxis to host plants to improve crop yield and protect watersheds.Master of ScienceIncreasing awareness of global climate change has drastically focused attention on finding solutions to reduce environmental impacts while still providing sufficient food for the increasing world population. Beneficial Nitrogen Fixing (BNF) microbes may offer a feasible solution in reducing environmental impacts by supplying biological nitrogen to plants, which reduces fertilizer use and ultimately runoff. One-well studied beneficial microbial model is that of Sinorhizobium meliloti and its legume host Medicago sativa (alfalfa), the fourth most cultivated USA crop used for animal feed. Advancing research for this symbiosis model can provide solutions to enhance crop yield while diminishing environmental impacts
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