Progress 10/01/11 to 09/30/15
Outputs
Target Audience:This project addresses a fundamental question: how do bacteria sense surfaces and how do the properties of surfaces influence cell physiology. We anticipate that the results of this research will lead to applications and technology that protects the health of humans in Wisconsin. There is a strong connection to human health through the design of mechanisms for preventing infections. Similarly, a strong mechanism connects this research to the health of livestock in Wisconsin and to the protection of food products derived from these animals, which is prone to contamination and the growth of bacteria. We anticipate that the results of these studies will provide mechanisms of treating and preventing infections. Our target audience for this fundamental research are chemists, engineers, and biologists working to study microorganisms; the target audience for the applied aspect of this research are people and animals in Wisconsin. Since microbes have no geographic boundaries, we anticipate that this research extends beyond Wisconsin to impact Americans living in other states. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?The research provided training to several graduate students (including Thiago Santos, who is an under-represented minority), postdoctoral fellows, undergraduate students, local teachers (through summer internships), and local K-12 students (by working with their teachers and developing educational materials). How have the results been disseminated to communities of interest?The results have been disseminated through a number of different mechanisms: 1. Publications in scientific journals 2. Posters and presentations at conferences and at international universities 3. Science outreach activities at UW-Madison 4. K-12 classrooms 5. Educational magazines and newsletters What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
The results of the proposed research are providing insights into the role of physical interactions in the coordination of cell behavior and physiology that is relevant to the health of humans and livestock, and is directly connected to protecting the health of people and animals in Wisconsin. It may also uncover mechanisms of extracellular sensing that are widely conserved in Eubacteria and have application to other microbes that affect agriculture and livestock in Wisconsin. This knowledge will expand our understanding of the human microbiome and decipher mechanisms that are involved in the colonization of tissues by bacteria, which may lead to life threatening infections if untreated. The differentiation of cells is accompanied by increased tolerance to antibiotics. An understanding of the molecular mechanisms involved in this process may provide targets for the development of new antimicrobials, which could be used to alleviate the development of resistance to our most clinically important antibiotics. Our research focused on three aims summarized for which the outcomes are summarized below.To study the stimuli, sensor, and role of adhesion in Escherichia coli cell and Proteus mirabilis cell differentiation, we are using a multidisciplinary approach that combines polymer chemistry, biophysics, genomics, and single-cell microbiology. The proposed research hinges on three specific aims. In Aim 1, we worked on determining and characterizing the stimulus that triggers E. coli and P. mirabilis cell differentiation on polymer surfaces. To study this question, we synthesized hydrogel and non-hydrogel polymers with defined physical and chemical properties and used flow cytometry and microscopy to quantify the response of cells to different chemical and physical properties of surfaces. This approach is essentially new to the field of microbiology (in which the vast majority of polymers used are heterogeneous and isolated from natural materials) and made it possible for us to quantitatively study the stimuli of surfaces that cells sense and respond to. In Aim 2, we determined the role of adhesion in the differentiation of E. coli and P. mirabilis cells on surfaces. Our preliminary data suggests that type 1 pili are important organelles in the response of bacteria to surfaces and that sensing may be dependent on the adhesion of cells to surfaces and the stiffness of the surface. Our hypothesis is that cells adhere to surface using pili, cell growth distributes a force across the substrate (transmitted through pili), and that surface elasticity places a resistive load on the cell wall that may be transmitted into cells by surface-associated proteins, such as mechanosensitive ion channels. To quantify the role of pili in surface sensing, we synthesized polymer surfaces with defined chemical and physical properties and measured the cellular response to these surfaces using the techniques described in Aim 1. We correlated this response to adhesion by type 1 pili using quartz crystal microbalance measurements with dissipation monitoring to measure adhesion force between cells and polymer surfaces. This data enabled us to understand how adhesion is connected to surface sensing. This quantitative approach hinges on the fusion of techniques from chemistry, materials science, biophysics, and microbiology and is orthogonal to the direction of other approaches in the field. In Aim 3, we determined the E. coli and P. mirabilis sensor(s) that senses the extracellular environment and triggers differentiation, and the downstream biochemical and transcriptional network. This enabled us to identify a candidate list of genes for further studies. In the future, we will use other -omics techniques to continue these studies. An exciting direction that emerged during these studies was the behavior of bacteria in liquid-crystalline environments and how these anisotropic materials alter cell behavior and motility. In essence, we founded an entirely new area of cell motility and have been focused on expanding this area rapidly.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
R. R. Trivedi, R. Maeda, S. E. Spagnolie, N. L. Abbott, D. B. Weibel. Bacterial Transport of Colloids in Liquid Crystalline Environments, 2015, Soft Matter 11, 8404-8408.
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
11. P. C. Mushenheim, R. R. Trivedi, S. S. Roy, M. S. Arnold, D. B. Weibel, N. L. Abbott. Effects of Confinement, Surface-Induced Orientations and Strain on Dynamical Behaviors of Bacteria in Thin Liquid Crystalline Films, 2015, Soft Matter 11, 6821-6831.
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
T.Y. Lin, T.M.A. Santos, W.S. Kontur, T.J. Donohue, D.B. Weibel. A cardiolipin-deficient mutant of Rhodobacter sphaeroides has an altered cell shape and is impaired in biofilm formation, Journal of Bacteriology, 2015, 197, 3446-3455.
- Type:
Journal Articles
Status:
Published
Year Published:
2015
Citation:
N. Yin, M.D. Stilwell, T.M.A. Santos, H. Wang, D.B. Weibel. Agarose particle-templated porous bacterial cellulose and its application in cartilage growth in vitro, Acta Biomateriala, 2015, 12, 129-138.
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Progress 10/01/13 to 09/30/14
Outputs
Target Audience: The target audiences are: undergraduates and graduate students, postdoctoral fellows, K-12 students and teachers, and local families. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? The research has provided training to several graduate students (including Thiago Santos, who is an under-represented minority), postdoctoral fellows, undergraduate students, local teachers (through summer internships), and local K-12 students, by working with their teachers and developing educational materials. How have the results been disseminated to communities of interest? The results have been disseminated through a number of different mechanisms: 1. Publications in scientific journals 2. Posters and presentations at conferences and international universities 3. Science outreach activities at the University of Wisconsin-Madison 4. K-12 classrooms 5. Educational magazines and newsletters What do you plan to do during the next reporting period to accomplish the goals? We plan to continue to work on the 3 aims identified below: To study the stimuli, sensor, and role of adhesion in E. coli cell and P. mirabilis cell differentiation, we are using a multidisciplinary approach that combines polymer chemistry, biophysics, genomics, and single-cell microbiology. The proposed research hinges on three specific aims. In Aim 1, we are determining and characterizing the stimulus that triggers E. coli and P. mirabilis cell differentiation on polymer surfaces. To study this question, we are synthesizing hydrogel and non-hydrogel polymers with defined physical and chemical properties and will use flow cytometry and microscopy to quantify the response of cells to different chemical and physical properties of surfaces. This approach is essentially new to the field of microbiology (in which the vast majority of polymers used are heterogeneous and isolated from natural materials) and is making it possible for us to quantitatively study the stimuli of surfaces that cells sense and respond to. In Aim 2, we are determining the role of adhesion in the differentiation of E. coli and P. mirabilis cells on surfaces. Our preliminary data suggests that type 1 pili are important organelles in the response of bacteria to surfaces and that sensing may be dependent on the adhesion of cells to surfaces and the stiffness of the surface. Our hypothesis is that cells adhere to surface using pili, cell growth distributes a force across the substrate (transmitted through pili), and that surface elasticity places a resistive load on the cell wall that may be transmitted into cells by surface-associated proteins, such as mechanosensitive ion channels. To quantify the role of pili in surface sensing, we are synthesizing polymer surfaces with defined chemical and physical properties and are measuring the cellular response to these surfaces using the techniques described in Aim 1. We are correlating this response to adhesion by type 1 pili using quartz crystal microbalance measurements with dissipation monitoring to measure adhesion force between cells and polymer surfaces. The analysis of this data is enabling us to understand how adhesion is connected to surface sensing. This quantitative approach hinges on the fusion of techniques from chemistry, materials science, biophysics, and microbiology and is orthogonal to the direction of other approaches in the field. In Aim 3, we are determining the E. coli and P. mirabilis sensor(s) that senses the extracellular environment and triggers differentiation, and the downstream biochemical and transcriptional network. We will incubate bacterial cells on polymer surfaces on which we know that cells respond (Aims 1 and 2), will harvest these cells, isolate their mRNA, and use genomic approaches to identify up/down-regulated transcripts. This approach will enable us to identify a candidate list of genes for further studies. Other -omics techniques will be used as necessary.
Impacts
What was accomplished under these goals?
The results of the proposed research are providing insights into the role of physical interactions in the coordination of cell behavior and physiology that is relevant to the health of humans and livestock, which is directly connected to protecting the health of people and animals in Wisconsin. It may also uncover mechanisms of extracellular sensing that are widely conserved in Eubacteria and have application to other microbes that affect agriculture and livestock in Wisconsin. This knowledge will expand our understanding of the human microbiome and decipher mechanisms that are involved in the colonization of tissues by bacteria, which may lead to life threatening infections if untreated. The differentiation of cells is accompanied by increased tolerance to antibiotics. An understanding of the molecular mechanisms involved in this process may provide targets for the development of new antimicrobials, which could be used to alleviate the development of resistance to our most clinically important antibiotics.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
J. M. Swiecicki, O. Sliusarenko, D. B. Weibel. From Swimming to Swarming: Escherichia coli Motility in Two-Dimensions, Integrative Biology, 2013, 5, 1490-1494.
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
N. Yin, T. Santos, G. Auer, .Oliver, D. B. Weibel. Bacterial Cellulose as a Substrate for Bacterial Cell Culture, Applied and Environmental Microbiology, 2014, 80, 1926-1932.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
H. H. Tuson, D. B. Weibel. Bacteria-Surface Interactions, Soft Matter, 2013, 9, 4368-4380.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
H. H. Tuson, M. F. Copeland, S. Carey, R. Sacotte, D. B. Weibel. Flagella Density Regulates Proteus mirabilis Swarmer Cell Motility in Viscous Environments, Journal of Bacteriology, 2013, 195, 368-377.
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
P.C. Mushenheim, R.R. Trivedi, D.B. Weibel, N.L. Abbott. "Using liquid crystals to reveal how mechanical anisotropy changes interfacial behaviors of motile bacteria," Biophysical Journal, 2014, 107, 255-265.
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Progress 01/01/13 to 09/30/13
Outputs
Target Audience: The target audience of this research is Wisconsin residents. Additionally the results can be used to improve livestock and crop health in areas outside of Wisconsin, which will improve domestic human health. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Many. Participants of this project have the opportunity to develop leadership skills by learning how to lead scientific projects. Participants can mentor undergraduate students who participate in the project. Participants may attend domestic meetings to present data, which contributes to their scientific development and professional develop. Participants also work closely with me on reporting and writing manuscripts to improve their written communication skills, and give talks on campus and at domestic venues (on this research) to improve their verbal communication skills. How have the results been disseminated to communities of interest? Results have been disseminated through manuscripts and conference proceedings. Several of the projects have also been used for science outreach activities to improve science education in Wisconsin. What do you plan to do during the next reporting period to accomplish the goals? We will continue along the sharp trajectory that we are currently on. We have already accomplished several of the goals of the proposed project and will continue to follow out research plan, which has been successful to date.
Impacts
What was accomplished under these goals?
The human body contains a staggering number and diversity of bacteria that form multicellular communities. In these structures, a division of labor protects bacterial cells against environmental stress and preserves the genotype of the parent strain. The goal of the proposed research is to build quantitative relationships between environmental cues and cells and is designed to probe how surfaces regulate and reprogram bacterial physiology. As model systems, our research focuses on the human intestinal bacterium Escherichia coli and the opportunistic urinary pathogen Proteus mirabilis. Both organisms play a role in the health of humans and livestock. Physical mechanisms are involved in controlling population-wide behavior and physiology in E. coli and P. mirabilis communities. We will address how the adsorption of E. coli and P. mirabilis at physical interfaces influences cell physiology by promoting the morphological differentiation of cells. The process of differentiation is directly linked to the attenuation of pathogenicity of E. coli and P. mirabilis and is an early step in the formation of pathogenic communities. To study the stimuli, sensor, and role of adhesion in E. coli cell and P. mirabilis cell differentiation, we are using a multidisciplinary approach that combines polymer chemistry, biophysics, genomics, and single-cell microbiology. The proposed research hinges on three specific aims. In Aim 1, we are determining and characterizing the stimulus that triggers E. coli and P. mirabilis cell differentiation on polymer surfaces. To study this question, we are synthesizing hydrogel and non-hydrogel polymers with defined physical and chemical properties and will use flow cytometry and microscopy to quantify the response of cells to different chemical and physical properties of surfaces. This approach is essentially new to the field of microbiology (in which the vast majority of polymers used are heterogeneous and isolated from natural materials) and is making it possible for us to quantitatively study the stimuli of surfaces that cells sense and respond to. In Aim 2, we are determining the role of adhesion in the differentiation of E. coli and P. mirabilis cells on surfaces. Our preliminary data suggests that type 1 pili are important organelles in the response of bacteria to surfaces and that sensing may be dependent on the adhesion of cells to surfaces and the stiffness of the surface. Our hypothesis is that cells adhere to surface using pili, cell growth distributes a force across the substrate (transmitted through pili), and that surface elasticity places a resistive load on the cell wall that may be transmitted into cells by surface-associated proteins, such as mechanosensitive ion channels. To quantify the role of pili in surface sensing, we are synthesizing polymer surfaces with defined chemical and physical properties and are measuring the cellular response to these surfaces using the techniques described in Aim 1. We are correlate this response to adhesion by type 1 pili using quartz crystal microbalance measurements with dissipation monitoring to measure adhesion force between cells and polymer surfaces. The analysis of this data is enabling to understand how adhesion is connected to surface sensing. This quantitative approach hinges on the fusion of techniques from chemistry, materials science, biophysics, and microbiology and is orthogonal to the direction of other approaches in the field. In Aim 3 we are determining the E. coli and P. mirabilis sensor(s) that senses the extracellular environment and triggers differentiation, and the downstream biochemical and transcriptional network. We will incubate bacterial cells on polymer surfaces on which we know that cells respond (Aims 1 and 2), will harvest these cells, isolate their mRNA, and use genomic approaches to identify up/down-regulated transcripts. This approach will enable us to identify a candidate list of genes for further studies. Other -omics techniques will be used as necessary.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2012
Citation:
J. M. Swiecicki, O. Sliusarenko, D. B. Weibel. From Swimming to Swarming: Escherichia coli Motility in Two-Dimensions, 2013, Integrative Biology, 5, 1490-1494.
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
N. Yin, T. Santos, D. B. Weibel. Bacterial Cellulose as a Substrate for Bacterial Cell Culture, 2014, Applied and Environmental Microbiology, in press.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
H. H. Tuson, D. B. Weibel. Bacteria-Surface Interactions, Soft Matter 2013, 9, 4368-4380.
- Type:
Journal Articles
Status:
Published
Year Published:
2013
Citation:
H. H. Tuson, M. F. Copeland, S. Carey,** R. Sacotte,** D. B. Weibel. Flagella Density Regulates Proteus mirabilis Swarmer Cell Motility in Viscous Environments, Journal of Bacteriology 2013, 195, 368-377.
- Type:
Journal Articles
Status:
Published
Year Published:
2012
Citation:
H. H. Tuson, L. D. Renner, D. B. Weibel. Polyacrylamide Hydrogels as Substrates for Studying Bacteria, Chemical Communications 2012, 48, 1595-1597.
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Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: The human body contains a staggering number and diversity of bacteria that form multicellular communities. In these structures, a division of labor protects bacterial cells against environmental stress and preserves the genotype of the parent strain. The goal of the proposed research is to build quantitative relationships between environmental cues and cells and is designed to probe how surfaces regulate and reprogram bacterial physiology. As a model system, our research focuses on the human intestinal bacterium Escherichia coli. Physical mechanisms are involved in controlling population-wide behavior and physiology in E. coli communities. We will address how the adsorption of E. coli at physical interfaces influences cell physiology by promoting the morphological differentiation of cells. The process of differentiation is directly linked to the attenuation of pathogenicity in E. coli and is an early step in the formation of pathogenic communities. To study the stimuli, sensor, and role of adhesion in E. coli cell differentiation, we propose a multidisciplinary approach that combines polymer chemistry, biophysics, genomics, and single-cell microbiology. The proposed research hinges on three specific aims. In Aim 1, we will determine and characterize the stimulus that triggers E. coli cell differentiation on polymer surfaces. To study this question, we will synthesize hydrogel polymers with defined physical and chemical properties and will use flow cytometry and microscopy to quantify the response of cells to different chemical and physical properties of surfaces. These approaches are new to the field and will make it possible for us to quantitatively study the stimuli of surfaces that cells sense and respond to. In Aim 2, we will determine the role of adhesion in the differentiation of E. coli cells on surfaces. Our preliminary data suggests that type 1 pili are important in the response of bacteria to surfaces. To quantify their role in surface sensing, we will synthesize polymer surfaces with defined properties and will measure the cellular response to these surfaces using the techniques described in Aim 1. We will then correlate this response to adhesion by type 1 pili using quartz crystal microbalance measurements with dissipation monitoring to measure adhesion force between cells and polymer surfaces. The analysis of this data will enable us in understanding how adhesion is connected to surface sensing. This quantitative approach hinges on the fusion of techniques from chemistry, materials science, biophysics, and microbiology and is orthogonal to the direction of other approaches in the field. In Aim 3, we will determine the E. coli sensor that senses the extracellular environment and triggers differentiation, and the downstream biochemical and transcriptional network. We will incubate bacterial cells on polymer surfaces on which we know that cells respond (Aims 1 and 2), will harvest these cells, isolate their mRNA, and will use gene chips to identify up/down regulated transcripts. This approach will enable us to identify a candidate list of genes for further studies. Other -omics techniques will be used as necessary. PARTICIPANTS: Hannah Tuson, Matt Copeland, Douglas Weibel TARGET AUDIENCES: No changes. PROJECT MODIFICATIONS: No changes.
Impacts
The results of the proposed research will provide insights into the role of physical interactions in the coordination of cell behavior and physiology that is relevant to the health of humans and livestock. It may also uncover mechanisms of extracellular sensing that are widely conserved in Eubacteria and have application to other microbes that affect agriculture and livestock in Wisconsin. This knowledge will expand our understanding of the human microbiome and decipher mechanisms that are involved in the colonization of tissues by bacteria, which may lead to life threatening infections if untreated. The differentiation of cells is accompanied by increased tolerance to antibiotics. An understanding of the molecular mechanisms involved in this process may provide targets for the development of new antimicrobials, which could be used to alleviate the development of resistance to our most clinically important antibiotics.
Publications
- H. H. Tuson, M. F. Copeland, S. Carey, R. Sacotte, D. B. Weibel (2013) Flagella Density Regulates Proteus mirabilis Swarmer Cell Motility in Viscous Environments, Journal of Bacteriology, 195, 368-377.
- H H. Tuson, L. D. Renner, D. B. Weibel (2012) Polyacrylamide Hydrogels as Substrates for Studying Bacteria, Chemical Communications, 48, 1595-1597.
- H. H. Tuson, D.B. Weibel (2013) Bacteria-Surface Interactions, manuscript submitted.
- J. M. Swiecicki, O. Sliusarenko, D. B. Weibel (2013) From Swimming to Swarming: Escherichia coli Motility in Two-Dimensions, manuscript submitted.
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: This is a new project that started in October, 2011, and hence there is limited data to report to-date. The project output of this project will lead to several new collaborations--including a new international collaboration that we are now trying to establish with Prof. Tilo Pompe in the Department of Bioengineering, University of Leipzig--as well as others. The output will include training at least one graduate student (Hannah Tuson), at least one postdoctoral fellow (Lars Renner), several undergraduate students (including Hannah Sandock and Anyi Wang), and potentially several high school students (Norah Ntambi, John Ntambi). PARTICIPANTS: Postdoctoral fellow: Lars Renner; Graduate students: Hannah Tuson; others as the project progresses. Undergraduate students: Hannah Sandock; others as the project progresses High School students: Norah Ntambi TARGET AUDIENCES: University of students; K-12 students; parents; local teachers PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
The results of this research are poised to extend our understanding of how bacterial sense surfaces and interfaces and how these interactions reprogram transcription of the genome and the biochemistry of cells. The outcome will improve our systems level understanding of bacteria as well have having long-term applications to the treatment of infections and the suppression of bacterial pathogenesis. The techniques and data may also shed light on the culture of new bacteria that have applications to food safety and livestock and plant culture in Wisconsin.
Publications
- No publications reported this period
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