Mechanism and Function of Membrane Homeostasis of Sortase Modulated by an Evolutionarily Conserved Protein Involved in Pilus Assembly in Actinobacteria PDF Download
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Author: Nicholas Anthony Ramirez
Publisher:
ISBN:
Category :
Languages : en
Pages : 99
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Book Description
Bacteria utilize proteins at their surface for a multitude of processes including adhesion, biofilm formation, motility, and virulence. Thus, understanding the biogenesis and surface display of these factors is instrumental in our understanding of bacterial pathogenesis and virulence mechanisms. Within this dissertation we describe the identification and characterization of a newly identified peptide which is functionally conserved amongst Actinobacteria and serves to modulate anchoring of proteins to the cell wall through modulation of membrane homeostasis of the housekeeping sortase. In the oral cavity associated bacterial species, Actinomyces oris, we identified a small peptide consisting of 52 amino acids which is encoded directly downstream of the gene encoding the housekeeping sortase SrtA. Henceforth we refer to this peptide as SafA for Sortase Associated Factor A. Firstly, through bioinformatic analysis we found that nearly all Actinobacteria encode a SafA homolog immediately downstream of their respective housekeeping sortase genes, with the exception of Bifidobacterium dentium in which the genome does not contain a separate SafA reading frame, but rather the C-terminus of the housekeeping sortase harbors a domain homologous to SafA in A. oris. In A. oris we found that deletion of safA results in phenotypes consistent with deletion of the housekeeping sortase itself, which include the formation of abnormally long pili as detected by electron microscopy and the failure of A. oris to interact with another oral bacterial species Streptococcus oralis. Cellular fractionation and immunoblotting revealed that in the absence of SafA, SrtA is cleaved and released into the extracellular milieu. While software predictions did not identify a signal peptide sequence in SrtA, manual amino acid sequence, sequence analysis did in fact reveal that SrtA contains a tripartite domain consistent with a type I signal peptide sequence and a predicted cleavage site between A56 and S57. Edman degradation amino acid sequencing confirmed this cleavage site and mutational analysis revealed that the signal peptidase LepB2 is responsible for this observed cleavage of SrtA. To elucidate how SafA protects SrtA from cleavage we utilized a Bacterial Adenylate Cyclase Two-Hybrid system which demonstrated that SafA and SrtA directly interact. Furthermore, we identified a three amino acid domain in SafA consisting of FPW residues which is essential for mediating this interaction. Finally, we found that ectopic expression of SafA from A. oris, Corynebacterium diphtheriae, and Corynebacterium matruchotii rescued the aforementioned functional defects of the safA mutant of A. oris, thus supporting the conclusion that SafA is both functionally and evolutionarily conserved. The findings described herein demonstrate a new paradigm for the modulation surface protein display in Actinobacteria. The conservation of SafA across Actinobacteria coupled with the essential role for sortases in mediating anchoring of pili and key virulence factors provides a unique target and opportunity to inhibit the virulence of Actinobacteria species.
Author: Nicholas Anthony Ramirez
Publisher:
ISBN:
Category :
Languages : en
Pages : 99
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Book Description
Bacteria utilize proteins at their surface for a multitude of processes including adhesion, biofilm formation, motility, and virulence. Thus, understanding the biogenesis and surface display of these factors is instrumental in our understanding of bacterial pathogenesis and virulence mechanisms. Within this dissertation we describe the identification and characterization of a newly identified peptide which is functionally conserved amongst Actinobacteria and serves to modulate anchoring of proteins to the cell wall through modulation of membrane homeostasis of the housekeeping sortase. In the oral cavity associated bacterial species, Actinomyces oris, we identified a small peptide consisting of 52 amino acids which is encoded directly downstream of the gene encoding the housekeeping sortase SrtA. Henceforth we refer to this peptide as SafA for Sortase Associated Factor A. Firstly, through bioinformatic analysis we found that nearly all Actinobacteria encode a SafA homolog immediately downstream of their respective housekeeping sortase genes, with the exception of Bifidobacterium dentium in which the genome does not contain a separate SafA reading frame, but rather the C-terminus of the housekeeping sortase harbors a domain homologous to SafA in A. oris. In A. oris we found that deletion of safA results in phenotypes consistent with deletion of the housekeeping sortase itself, which include the formation of abnormally long pili as detected by electron microscopy and the failure of A. oris to interact with another oral bacterial species Streptococcus oralis. Cellular fractionation and immunoblotting revealed that in the absence of SafA, SrtA is cleaved and released into the extracellular milieu. While software predictions did not identify a signal peptide sequence in SrtA, manual amino acid sequence, sequence analysis did in fact reveal that SrtA contains a tripartite domain consistent with a type I signal peptide sequence and a predicted cleavage site between A56 and S57. Edman degradation amino acid sequencing confirmed this cleavage site and mutational analysis revealed that the signal peptidase LepB2 is responsible for this observed cleavage of SrtA. To elucidate how SafA protects SrtA from cleavage we utilized a Bacterial Adenylate Cyclase Two-Hybrid system which demonstrated that SafA and SrtA directly interact. Furthermore, we identified a three amino acid domain in SafA consisting of FPW residues which is essential for mediating this interaction. Finally, we found that ectopic expression of SafA from A. oris, Corynebacterium diphtheriae, and Corynebacterium matruchotii rescued the aforementioned functional defects of the safA mutant of A. oris, thus supporting the conclusion that SafA is both functionally and evolutionarily conserved. The findings described herein demonstrate a new paradigm for the modulation surface protein display in Actinobacteria. The conservation of SafA across Actinobacteria coupled with the essential role for sortases in mediating anchoring of pili and key virulence factors provides a unique target and opportunity to inhibit the virulence of Actinobacteria species.
Author: Hans-Curt Flemming
Publisher: IWA Publishing
ISBN: 1780407416
Category : Science
Languages : en
Pages : 336
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Book Description
The Perfect Slime presents the latest state of knowledge and all aspects of the Extracellular Polymeric Substances, (EPS) matrix – from the ecological and health to the antifouling perspectives. The book brings together all the current material in order to expand our understanding of the functions, properties and characteristics of the matrix as well as the possibilities to strengthen or weaken it. The EPS matrix represents the immediate environment in which biofilm organisms live. From their point of view, this matrix has paramount advantages. It allows them to stay together for extended periods and form synergistic microconsortia, it retains extracellular enzymes and turns the matrix into an external digestion system and it is a universal recycling yard, it protects them against desiccation, it allows for intense communication and represents a huge genetic archive. They can remodel their matrix, break free and eventually, they can use it as a nutrient source. The EPS matrix can be considered as one of the emergent properties of biofilms and are a major reason for the success of this form of life. Nevertheless, they have been termed the “black matter of biofilms” for good reasons. First of all: the isolation methods define the results. In most cases, only water soluble EPS components are investigated; insoluble ones such as cellulose or amyloids are much less included. In particular in environmental biofilms with many species, it is difficult to impossible isolate, separate the various EPS molecules they are encased in and to define which species produced which EPS. The regulation and the factors which trigger or inhibit EPS production are still very poorly understood. Furthermore: bacteria are not the only microorganisms to produce EPS. Archaea, Fungi and algae can also form EPS. This book investigates the questions, What is their composition, function, dynamics and regulation? What do they all have in common?
Author: Ephraim Cohen
Publisher: Springer
ISBN: 3030129195
Category : Science
Languages : en
Pages : 820
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Book Description
The extracellular matrix (ECM) is an acellular three-dimensional network composed of proteins, glycoproteins, proteoglycans and exopolysaccharides. It primarily serves as a structural component in the tissues and organs of plants and animals, or forms biofilms in which bacterial cells are embedded. ECMs are highly dynamic structures that undergo continuous remodeling, and disruptions are frequently the result of pathological processes associated with severe diseases such as arteriosclerosis, neurodegenerative illness or cancer. In turn, bacterial biofilms are a source of concern for human health, as they are associated with resistance to antibiotics. Although exopolysaccharides are crucial for ECM formation and function, they have received considerably little attention to date. The respective chapters of this book comprehensively address such issues, and provide reviews on the structural, biochemical, molecular and biophysical properties of exopolysaccharides. These components are abundantly produced by virtually all taxa including bacteria, algae, plants, fungi, invertebrates and vertebrates. They include long unbranched homopolymers (cellulose, chitin/chitosan), linear copolymers (alginate, agarose), peptoglycans such as murein, heteropolymers like a variety of glycosaminoglycans (hyaluronan, dermatan, keratin, heparin, Pel), and branched heteropolymers such as pectin and hemicellulose. A separate chapter is dedicated to modern industrial and biomedical applications of exopolysaccharides and polysaccharide-based biocomposites. Their unique chemical, physical and mechanical properties have attracted considerable interest, inspired basic and applied research, and have already been harnessed to form structural biocomposite hybrids for tailor-made applications in regenerative medicine, bioengineering and biosensor design. Given its scope, this book provides a substantial source of basic and applied information for a wide range of scientists, as well as valuable textbook for graduate and advanced undergraduate students.
Author: Pallaval Veera Bramhachari
Publisher: Springer
ISBN: 9811324298
Category : Medical
Languages : en
Pages : 381
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Book Description
This book illustrates the importance and significance of Quorum sensing (QS), it’s critical roles in regulating diverse cellular functions in microbes, including bioluminescence, virulence, pathogenesis, gene expression, biofilm formation and antibiotic resistance. Microbes can coordinate population behavior with small molecules called autoinducers (AHL) which serves as a signal of cellular population density, triggering new patterns of gene expression for mounting virulence and pathogenesis. Therefore, these microbes have the competence to coordinate and regulate explicit sets of genes by sensing and communicating amongst themselves utilizing variety of signals. This book descry emphasizes on how bacteria can coordinate an activity and synchronize their response to external signals and regulate gene expression. The chapters of the book provide the recent advancements on various functional aspects of QS systems in different gram positive and gram negative organisms. Finally, the book also elucidates a comprehensive yet a representative description of a large number of challenges associated with quorum sensing signal molecules viz. virulence, pathogenesis, antibiotic synthesis, biosurfactants production, persister cells, cell signaling and biofilms, intra and inter-species communications, host-pathogen interactions, social interactions & swarming migration in biofilms.
Author: Byung Hong Kim
Publisher: Cambridge University Press
ISBN: 1107171733
Category : Medical
Languages : en
Pages : 509
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Book Description
Extensive and up-to-date review of key metabolic processes in bacteria and archaea and how metabolism is regulated under various conditions.
Author: Byung Hong Kim
Publisher: Cambridge University Press
ISBN: 113946762X
Category : Science
Languages : en
Pages : 934
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Book Description
Recent determination of genome sequences for a wide range of bacteria has made in-depth knowledge of prokaryotic metabolic function essential in order to give biochemical, physiological, and ecological meaning to the genomic information. Clearly describing the important metabolic processes that occur in prokaryotes under different conditions and in different environments, this advanced text provides an overview of the key cellular processes that determine bacterial roles in the environment, biotechnology, and human health. Prokaryotic structure is described as well as the means by which nutrients are transported into cells across membranes. Glucose metabolism through glycolysis and the TCA cycle are discussed, as well as other trophic variations found in prokaryotes, including the use of organic compounds, anaerobic fermentation, anaerobic respiratory processes, and photosynthesis. The regulation of metabolism through control of gene expression and control of the activity of enzymes is also covered, as well as survival mechanisms used under starvation conditions.
Author: Otto Geiger
Publisher: Springer
ISBN: 9783319504292
Category : Science
Languages : en
Pages : 0
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Book Description
Concise chapters, written by experts in the field, cover a wide spectrum of topics on lipid and membrane formation in microbes (Archaea, Bacteria, eukaryotic microbes).All cells are delimited by a lipid membrane, which provides a crucial boundary in any known form of life. Readers will discover significant chapters on microbial lipid-carrying biomolecules and lipid/membrane-associated structures and processes.
Author: Raghvendra Pratap Singh
Publisher: Springer Nature
ISBN: 9811530289
Category : Science
Languages : en
Pages : 227
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Book Description
The book compiles the latest studies on microorganisms thriving in extreme conditions. Microbes have been found in extremely high and low temperatures, highly acidic to saline conditions, from deserts to the Dead sea, from hot-springs to underwater hydrothermal vents- the diversity is incredible. The various chapters highlight the microbial life and describe the mechanisms of tolerance to these harsh conditions, and show how an understanding of these phenomena can help us exploit the microbes in biotechnology. The theme of the book is highly significant since life in these environments can give vital clues about the origin and evolution of life on earth, as a lot of these conditions simulate the environment present billions of years ago. Additionally, the study of adaptation and survival of organisms in such environments can be important for finding life on other planets. This book shall be useful for students, researchers and course instructors interested in evolution, microbial adaptations and ecology in varied environments.
Author: Busi Siddhardha
Publisher: Springer Nature
ISBN: 9811516952
Category : Science
Languages : en
Pages : 685
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Book Description
This book provides essential insights into microbial pathogenesis, host-pathogen interactions, and the anti-microbial drug resistance of various human pathogens on the basis of various model organisms. The initial sections of the book introduce readers to the mechanisms of microbial pathogenesis, host-pathogen interactions, anti-microbial drug resistance, and the dynamics of biofilm formation. Due to the emergence of various microbial resistant strains, it is especially important to understand the prognosis for microbial infections, disease progression profiles, and mechanisms of resistance to antibiotic therapy in order to develop novel therapeutic strategies. In turn, the second part of the book presents a comparative analysis of various animal models to help readers understand microbial pathogenesis, host-pathogen interactions, anti-microbial drug discovery, anti-biofilm therapeutics, and treatment regimes. Given its scope, the book represents a valuable asset for microbiologists, biotechnologists, medical professionals, drug development researchers, and pharmacologists alike.
Author: Sunil Kumar
Publisher: Springer Nature
ISBN: 3030307573
Category : Medical
Languages : en
Pages : 318
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Book Description
This book highlights treatment strategies for bacterial biofilms in connection with a variety of human diseases. In particular, it reviews bacterial biofilm formation and its mechanism. Topics covered include biofilms in human health, the role of biofilms in mediating human diseases, and methods for testing bacterial biofilms. Further sections concentrate on biofilm-mediated diseases in different parts of the human gastrointestinal tract, while therapeutic strategies for biofilm control and natural agents that disrupt bacterial biofilms are also covered. Readers will also find the latest advances in probiotics and biofilms, as well as the use of probiotics to counteract biofilm-associated infections. Biofilms and antimicrobial resistance are discussed. Subsequent chapters address the management of inflammatory bowel disease via probiotics biofilms, as well as the role of probiotics bacteria in the treatment of human diseases associated with bacterial biofilms. The book is chiefly intended for clinicians/scientists in the fields of medical microbiology, applied microbiology, biochemistry, and biotechnology.
