Molecular Interactions: Oral Bacteria &; Matrix Proteins
Keith P Mintz, Research Associate Professor
Microbiology And Molecular Geneticsuniversity Of Vermont & St Agric College
Grant 5R01DE013824-09 from National Institute Of Dental & Craniofacial Research IRG: ODCS
Abstract: Actinobacillus actinomycetemcomitans is a Gram-negative, facultative anaerobic bacterium that colonizes the human oral cavity and the upper respiratory tract. This bacterium is strongly associated with localized aggressive periodontitis (LAP) and with cases of adult periodontitis. This pathogen is the causative agent for other serious infections including infectious endocarditis, soft tissue abscesses, pneumonia, and may contribute to cardiovascular disease. The periodontium is believed to be the source for these non-oral diseases, but little is known about the tropism used by A. actinomycetemcomitans to colonize the oral cavity and to infiltrate and disseminate in tissues. Pathogens have developed diverse strategies to be successful in colonization of host tissues. A common theme amongst these pathogens is the ability to initiate infection by adhesion to specific host macromolecules under stringent or hostile conditions. These molecules include proteins secreted by host cells that form the extracellular matrix (ECM). A. actinomycetemcomitans is found in the connective tissue of the periodontium and in close association with collagen fibers in infected tissues. The bacterium also binds to the ECM proteins, collagen, fibronectin and laminin. Using a genetic approach, we have identified the first A. actinomycetemcomitans collagen adhesin, Ema (extracellular matrix protein adhesin) A and multiple genes involved in regulating the expression of ECM protein adhesin activity. EmaA is structurally related to YadA, a multipurpose ECM protein adhesin of the enteropathogenic bacterium Yersinia enterocolitica, and is associated with bacterial cell surface appendages. These EmaA structures are proposed to be fundamental for collagen adhesion. To elucidate the role of EmaA in colonization and pathogenicity of the bacterium, we propose to 1) map the functional domains of EmaA by determining the collagen binding, subcellular localization, and assembly of surface structures in A. actinomycetemcomitans, 2) investigate the surface structures associated with EmaA by transmission electron microscopy and the role of these structures in resistance to the innate immune response of the host, and 3) determine the number of EmaA molecules required for the assembly of structures on the surface of A. actinomycetemcomitans. A long term goal of the proposed research is to identify and characterize bacterial adhesins that are required for the colonization of the oral cavity and non-oral tissues. These adhesins may serve as targets for future drug development involving small molecules or vaccines that disrupt host-pathogen interactions. Lay statement Binding to host tissues is the initial phase of all infectious diseases. Understanding how bacteria interact with host cells or tissue constituents will aid in the development of novel therapeutics to prevent initiation or progression of the disease
Project start date: 2000-07-01
Project end date: 2011-06-30
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Molecular Interactions: Oral Bacteria &; Matrix Proteins
Keith P Mintz, Research Associate Professor
Microbiology And Molecular Geneticsuniversity Of Vermont & St Agric College
85 South Prospect Street
burlington, Vt 05405
Grant 5R01DE013824-08 from National Institute Of Dental & Craniofacial Research IRG: ODCS
Abstract: Actinobacillus actinomycetemcomitans is a Gram-negative, facultative anaerobic bacterium that colonizes the human oral cavity and the upper respiratory tract. This bacterium is strongly associated with localized aggressive periodontitis (LAP) and with cases of adult periodontitis. This pathogen is the causative agent for other serious infections including infectious endocarditis, soft tissue abscesses, pneumonia, and may contribute to cardiovascular disease. The periodontium is believed to be the source for these non-oral diseases, but little is known about the tropism used by A. actinomycetemcomitans to colonize the oral cavity and to infiltrate and disseminate in tissues. Pathogens have developed diverse strategies to be successful in colonization of host tissues. A common theme amongst these pathogens is the ability to initiate infection by adhesion to specific host macromolecules under stringent or hostile conditions. These molecules include proteins secreted by host cells that form the extracellular matrix (ECM). A. actinomycetemcomitans is found in the connective tissue of the periodontium and in close association with collagen fibers in infected tissues. The bacterium also binds to the ECM proteins, collagen, fibronectin and laminin. Using a genetic approach, we have identified the first A. actinomycetemcomitans collagen adhesin, Ema (extracellular matrix protein adhesin) A and multiple genes involved in regulating the expression of ECM protein adhesin activity. EmaA is structurally related to YadA, a multipurpose ECM protein adhesin of the enteropathogenic bacterium Yersinia enterocolitica, and is associated with bacterial cell surface appendages. These EmaA structures are proposed to be fundamental for collagen adhesion. To elucidate the role of EmaA in colonization and pathogenicity of the bacterium, we propose to 1) map the functional domains of EmaA by determining the collagen binding, subcellular localization, and assembly of surface structures in A. actinomycetemcomitans, 2) investigate the surface structures associated with EmaA by transmission electron microscopy and the role of these structures in resistance to the innate immune response of the host, and 3) determine the number of EmaA molecules required for the assembly of structures on the surface of A. actinomycetemcomitans. A long term goal of the proposed research is to identify and characterize bacterial adhesins that are required for the colonization of the oral cavity and non-oral tissues. These adhesins may serve as targets for future drug development involving small molecules or vaccines that disrupt host-pathogen interactions. Lay statement Binding to host tissues is the initial phase of all infectious diseases. Understanding how bacteria interact with host cells or tissue constituents will aid in the development of novel therapeutics to prevent initiation or progression of the disease
Project start date: 2000-07-01
Project end date: 2011-06-30
5R01DE013824-08 (2008): $346676
Molecular Interactions: Oral Bacteria & Matrix Proteins
Keith P Mintz, Research Associate Professor
University Of Vermont And St Agric College 85 South Prospect Street Burlington, Vt 05405
Grant 5R01DE013824-07 from National Institute Of Dental And Craniofacial Research IRG: ODCS
Abstract: Actinobacillus actinomycetemcomitans is a Gram-negative, facultative anaerobic bacterium that colonizes the human oral cavity and the upper respiratory tract. This bacterium is strongly associated with localized aggressive periodontitis (LAP) and with cases of adult periodontitis. This pathogen is the causative agent for other serious infections including infectious endocarditis, soft tissue abscesses, pneumonia, and may contribute to cardiovascular disease. The periodontium is believed to be the source for these non-oral diseases, but little is known about the tropism used by A. actinomycetemcomitans to colonize the oral cavity and to infiltrate and disseminate in tissues. Pathogens have developed diverse strategies to be successful in colonization of host tissues. A common theme amongst these pathogens is the ability to initiate infection by adhesion to specific host macromolecules under stringent or hostile conditions. These molecules include proteins secreted by host cells that form the extracellular matrix (ECM). A. actinomycetemcomitans is found in the connective tissue of the periodontium and in close association with collagen fibers in infected tissues. The bacterium also binds to the ECM proteins, collagen, fibronectin and laminin. Using a genetic approach, we have identified the first A. actinomycetemcomitans collagen adhesin, Ema (extracellular matrix protein adhesin) A and multiple genes involved in regulating the expression of ECM protein adhesin activity. EmaA is structurally related to YadA, a multipurpose ECM protein adhesin of the enteropathogenic bacterium Yersinia enterocolitica, and is associated with bacterial cell surface appendages. These EmaA structures are proposed to be fundamental for collagen adhesion. To elucidate the role of EmaA in colonization and pathogenicity of the bacterium, we propose to 1) map the functional domains of EmaA by determining the collagen binding, subcellular localization, and assembly of surface structures in A. actinomycetemcomitans, 2) investigate the surface structures associated with EmaA by transmission electron microscopy and the role of these structures in resistance to the innate immune response of the host, and 3) determine the number of EmaA molecules required for the assembly of structures on the surface of A. actinomycetemcomitans. A long term goal of the proposed research is to identify and characterize bacterial adhesins that are required for the colonization of the oral cavity and non-oral tissues. These adhesins may serve as targets for future drug development involving small molecules or vaccines that disrupt host-pathogen interactions. Lay statement Binding to host tissues is the initial phase of all infectious diseases. Understanding how bacteria interact with host cells or tissue constituents will aid in the development of novel therapeutics to prevent initiation or progression of the disease.
Project start date: 2000-07-01
Project end date: 2011-06-30
5R01DE013824-07 (2007): $350532
5R01DE013824-05 (2005): $304000
Grants awarded to Keith P Mintz
Molecular Interactions: Oral Bacteria &; Matrix Proteins
Keith P Mintz, Research Assistant Professor
Microbiol & Molecular Geneticsuniversity Of Vermont & St Agric College
85 South Prospect Street
burlington, Vt 05405
Grant 1R01DE013824-01A1 from National Institute Of Dental & Craniofacial Research IRG: OBM
Abstract: Gram-negative bacteria are associated with periodontal diseases, which are a group of chronic inflammatory diseases of the gingiva and the supporting structures of the periodontium. Actinobacillus actinomycetemcomitans (Aa) is a Gram-negative, facultative coccobacillus that colonizes the human oral cavity and upper respiratory tract. This bacterium is closely associated with periodontitis in young individuals and with cases of adult periodontitis. This pathogen has been associated with other serious human infections such as endocarditis, soft tissue abscesses, and more recently cardiovascular disease. Although the periodontium is believed to be the source of these extraoral infections, little is known about the tropisms used by Aa to maintain itself within the oral cavity and to infiltrate and disseminate in tissues. Pathogens have evolved diverse strategies to be successful in colonization of the host tissue. A common theme amongst these pathogens is the ability to initiate infection by adhesion to specific host macromolecules under stringent or hostile conditions. These molecules include proteins that are secreted by host cells that form the extracellular matrix. This matrix is usually composed of collagen and specific noncollagenous proteins, e.g. fibronectin. The major protein found in the periodontium is collagen and we have demonstrated that Aa binds to both collagen and fibronectin. In this proposal, we plan to identify the genes coding for matrix binding proteins and determine the amino acid sequences of these proteins required for binding. It is our hypothesis that the synthesis of matrix binding proteins is involved in Aa colonization of the periodontal pocket and underlying tissues. In order to realize these goals, we propose to 1) isolate the genes coding for collagen and fibronectin binding proteins by constructing a transposon mutagenesis and a phage display library; 2) determine the gene sequences and generate isogenic mutants; 3) determine the regions(s) of the protein essential for binding activity; 4) determine the immunoreactivity of LJP patient sera to these proteins. This information can be used for the development of therapeutic agents of vaccines for periodontal disease
Keywords: Actinobacillus actinomycetemcomitans, Bacteroides gingivalis, bacteria infection mechanism, bacterial protein, binding protein, extracellular matrix protein, oral bacteria, periodontitis bacterial genetics, cell adhesion molecule, cell type, collagen, epithelium, fibronectin, genome, mutant, periodontium, protein sequence, protein structure, transposon /insertion element genetic library, human tissue, polymerase chain reaction, protein purification
Project start date: 2001-08-01
Project end date: 2004-06-30
1R01DE013824-01A1 (2001): $187475
Molecular Interactions: Oral Bacteria & Matrix Proteins
Keith P Mintz, Research Assistant Professor
University Of Vermont And St Agric College 85 South Prospect Street Burlington, Vt 05405
Grant 2R01DE013824-06 from National Institute Of Dental And Craniofacial Research IRG: ODCS
Abstract: Actinobacillus actinomycetemcomitans is a Gram-negative, facultative anaerobic bacterium that colonizes the human oral cavity and the upper respiratory tract. This bacterium is strongly associated with localized aggressive periodontitis (LAP) and with cases of adult periodontitis. This pathogen is the causative agent for other serious infections including infectious endocarditis, soft tissue abscesses, pneumonia, and may contribute to cardiovascular disease. The periodontium is believed to be the source for these non-oral diseases, but little is known about the tropism used by A. actinomycetemcomitans to colonize the oral cavity and to infiltrate and disseminate in tissues. Pathogens have developed diverse strategies to be successful in colonization of host tissues. A common theme amongst these pathogens is the ability to initiate infection by adhesion to specific host macromolecules under stringent or hostile conditions. These molecules include proteins secreted by host cells that form the extracellular matrix (ECM). A. actinomycetemcomitans is found in the connective tissue of the periodontium and in close association with collagen fibers in infected tissues. The bacterium also binds to the ECM proteins, collagen, fibronectin and laminin. Using a genetic approach, we have identified the first A. actinomycetemcomitans collagen adhesin, Ema (extracellular matrix protein adhesin) A and multiple genes involved in regulating the expression of ECM protein adhesin activity. EmaA is structurally related to YadA, a multipurpose ECM protein adhesin of the enteropathogenic bacterium Yersinia enterocolitica, and is associated with bacterial cell surface appendages. These EmaA structures are proposed to be fundamental for collagen adhesion. To elucidate the role of EmaA in colonization and pathogenicity of the bacterium, we propose to 1) map the functional domains of EmaA by determining the collagen binding, subcellular localization, and assembly of surface structures in A. actinomycetemcomitans, 2) investigate the surface structures associated with EmaA by transmission electron microscopy and the role of these structures in resistance to the innate immune response of the host, and 3) determine the number of EmaA molecules required for the assembly of structures on the surface of A. actinomycetemcomitans. A long term goal of the proposed research is to identify and characterize bacterial adhesins that are required for the colonization of the oral cavity and non-oral tissues. These adhesins may serve as targets for future drug development involving small molecules or vaccines that disrupt host-pathogen interactions. Lay statement Binding to host tissues is the initial phase of all infectious diseases. Understanding how bacteria interact with host cells or tissue constituents will aid in the development of novel therapeutics to prevent initiation or progression of the disease.
Keywords: adhesin, collagen, oral bacteria, protein, tissue, adhesion, extracellular matrix protein
Project start date: 2000-07-01
Project end date: 2011-06-30
2R01DE013824-06 (2006): $361000
2R01DE013824-04 (2004): $304000
Invasion And Adhesion Of Periopathogens
Keith P Mintz, Research Associate Professor
Microbiol & Molecular Geneticsuniversity Of Vermont & St Agric College
85 South Prospect Street
burlington, Vt 05405
Grant 5R01DE009760-15 from National Institute Of Dental & Craniofacial Research IRG: ZRG1
Keywords: adhesion, cell, microtubule Actinobacillus actinomycetemcomitans, Saccharomyces cerevisiae, antiserum, bacteria, biopsy, cell component, cell line, cytoplasm, cytoskeleton, element, endocarditis, gene, health /scientific organization, human, infection, inflammation, intracellular, larynx, library, literature survey, macromolecule, membrane, mutant, nose, nucleic acid sequence, organelle, pathology, periodontitis, periodontium, play, polymer, protein, quality of life, receptor, role, tissue, trachea, tubulin, yeast
Project start date: 1992-03-01
Project end date: 2008-11-30
5R01DE009760-15 (2007): $359123
5R01DE009760-14 (2006): $369849
BACTERIAL ADHESINS OF PERIOPATHOGENS
Keith P Mintz, Research Assistant Professor
University Of Vermont And St Agric College 85 South Prospect Street Burlington, Vt 05405
Grant 1F32DE005626-01 from National Institute Of Dental And Craniofacial Research IRG: DSR
1F32DE005626-01 (1992): $28600