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035 a| (Sirsi) o936213479;

035 a| (OCoLC)936213479;

049 a| EREE;

100 1 a| Betteken, Michael I.,e| author.?| UNAUTHORIZED;

245 10 a| Role of the iron storage ferritins Dps and DpsL during the prolonged oxidative stress response of Bacteroides fragilis /c| by Michael I. Betteken.;

264 1 a| [Greenville, N.C.] :b| [East Carolina University],c| 2015.;

300 a| 152 pages :b| illustrations, charts.;

336 a| textb| txt2| rdacontent;

337 a| computerb| c2| rdamedia;

338 a| online resourceb| cr2| rdacarrier;

347 a| text fileb| PDFc| 2.71 MB2| rda;

490 1 a| ECU Brody School of Medicine thesis;

538 a| System requirements: Adobe Reader.;

538 a| Mode of access: World Wide Web.;

502 b| Ph.D.c| East Carolina Universityd| 2015.;

500 a| Presented to the faculty of the Department of Microbiology and Immunology.;

500 a| Advisor: C. Jeffrey Smith.;

500 a| Title from PDF t.p. (viewed February 29, 2016).;

504 a| Includes bibliographical references.;

520 3 a| Bacteroides fragilis is a Gram negative anaerobe and member of the human intestinal tract microbiome. B. fragilis serves many beneficial roles within the intestinal tract; however, its translocation to the peritoneal cavity and the blood stream can result in peritonitis, intra-abdominal abscess formation, bacteremia and sepsis. We have shown that B. fragilis mediates both acute and Prolonged Oxidative Stress (POST) responses both in vitro and in vivo. This report characterizes the role and the genetic regulation of the iron storage proteins Dps and DpsL during the POST response. To test sensitivity to oxidative stress during the POST response a disk diffusion assay was developed using tert-butyl hydroperoxide (tBOOH). When the assay plates received aerobic exposure for three hours there was no zone of growth inhibition, whereas those kept under anaerobic conditions were highly sensitive to tBOOH. These results demonstrated an oxygen induced resistance to tBOOH that was mediated by prolonged aerobic exposure. To determine a mechanism for this POST induced resistance to tBOOH, a series of oxidative stress mutants were assayed. Only the [Delta]dps mutant was sensitive to tBOOH after aerobic exposure indicating that Dps mediated the POST phenotype. Because of the similarities to Dps, the recently characterized DpsL (bfr) was tested for a role in the POST response. The [Delta]bfr mutant demonstrated resistance to tBOOH after aerobic exposure similar to wildtype (WT); however, when a double [Delta]dps [Delta]bfr mutant was generated it demonstrated sensitivity to tBOOH that was greater than the [delta]dps mutant indicating that both Dps and DpsL play a role in the resistance phenotype. To explore the role that Dps and DpsL play in the survival of B. fragilis during infection, animal experiments were performed in the rat abscess model. Interestingly only the double [Delta]dps [Delta] mutant was attenuated in this model whereas neither of the single mutants showed a defect in competition experiments with WT. This indicated that both Dps and DpsL play a role in survival during infection. To investigate genetic regulation during the POST response, it was essential to identify the second regulator of dps expression. It was previously shown that OxyR is a strong inducer of dps expression during acute oxidative stress; however, the [Delta]oxyR mutant was resistant to tBOOH after prolonged aerobic exposure similar to WT. This indicated that there was a second regulator of dps expression during the POST response. A known POST regulator, SigOF was investigated to determine if it played a role in this response. Similar to [Delta]oxyR the [Delta]sigOF mutant was resistant to tBOOH. Interestingly though a double [Delta]sigOF [Delta]oxyR mutant was sensitive to tBOOH in the POST assay and dps expression was reduced as shown by qRTPCR. These results strongly suggest that SigOF is the regulator responsible for dps expression during the POST response.;

650 2 a| Oxidative Stress.=| ^A930446;

650 2 a| Bacteroides fragilis.=| ^A946076;

650 2 a| Ferritins.=| ^A1202664;

650 2 a| Oxidation-Reduction.=| ^A919450;

650 2 a| Physiological Processes.=| ^A1001681;

653 a| iron;

653 a| dps;

653 a| dpsL;

700 1 a| Smith, C. Jeffrey,e| degree supervisor.?| UNAUTHORIZED;

710 2 a| East Carolina University.b| Department of Microbiology and Immunology.?| UNAUTHORIZED;

830 0 a| ECU Brody School of Medicine thesis.=| ^A964744;

856 40 z| Access via ScholarShipu| http://hdl.handle.net/10342/5116;

949 o| wjh;

994 a| C0b| ERE;

096 a| QZ 180;

596 a| 1 4;

998 a| 4356303;

999 a| CLICK ON WEB ADDRESSw| ASISc| 1i| 4356303-1001l| JNETm| JOYNERr| Ys| Yt| JNE3ETDu| 1/29/2016x| ETDz| JERESOURCE;

999 a| CLICK ON WEB ADDRESSw| ASISc| 1i| 4356303-2001l| HSLELECm| HSLr| Ys| Yt| HEETDu| 1/29/2016x| ETDz| HERESOURCE;

Role of the iron storage ferritins Dps and DpsL during the prolonged oxidative stress response of Bacteroides fragilis / by Michael I. Betteken.

Author/creator	Betteken, Michael I. author.
Other author	Smith, C. Jeffrey, degree supervisor.
Other author	East Carolina University. Department of Microbiology and Immunology.
Format	Theses and dissertations
Publication	[Greenville, N.C.] : [East Carolina University], 2015.
Description	152 pages : illustrations, charts.
Supplemental Content	Access via ScholarShip
Subjects	Oxidative Stress. Bacteroides fragilis. Ferritins. Oxidation-Reduction. Physiological Processes.

Series	ECU Brody School of Medicine thesis ECU Brody School of Medicine thesis. ^A964744
Summary	Bacteroides fragilis is a Gram negative anaerobe and member of the human intestinal tract microbiome. B. fragilis serves many beneficial roles within the intestinal tract; however, its translocation to the peritoneal cavity and the blood stream can result in peritonitis, intra-abdominal abscess formation, bacteremia and sepsis. We have shown that B. fragilis mediates both acute and Prolonged Oxidative Stress (POST) responses both in vitro and in vivo. This report characterizes the role and the genetic regulation of the iron storage proteins Dps and DpsL during the POST response. To test sensitivity to oxidative stress during the POST response a disk diffusion assay was developed using tert-butyl hydroperoxide (tBOOH). When the assay plates received aerobic exposure for three hours there was no zone of growth inhibition, whereas those kept under anaerobic conditions were highly sensitive to tBOOH. These results demonstrated an oxygen induced resistance to tBOOH that was mediated by prolonged aerobic exposure. To determine a mechanism for this POST induced resistance to tBOOH, a series of oxidative stress mutants were assayed. Only the [Delta]dps mutant was sensitive to tBOOH after aerobic exposure indicating that Dps mediated the POST phenotype. Because of the similarities to Dps, the recently characterized DpsL (bfr) was tested for a role in the POST response. The [Delta]bfr mutant demonstrated resistance to tBOOH after aerobic exposure similar to wildtype (WT); however, when a double [Delta]dps [Delta]bfr mutant was generated it demonstrated sensitivity to tBOOH that was greater than the [delta]dps mutant indicating that both Dps and DpsL play a role in the resistance phenotype. To explore the role that Dps and DpsL play in the survival of B. fragilis during infection, animal experiments were performed in the rat abscess model. Interestingly only the double [Delta]dps [Delta] mutant was attenuated in this model whereas neither of the single mutants showed a defect in competition experiments with WT. This indicated that both Dps and DpsL play a role in survival during infection. To investigate genetic regulation during the POST response, it was essential to identify the second regulator of dps expression. It was previously shown that OxyR is a strong inducer of dps expression during acute oxidative stress; however, the [Delta]oxyR mutant was resistant to tBOOH after prolonged aerobic exposure similar to WT. This indicated that there was a second regulator of dps expression during the POST response. A known POST regulator, SigOF was investigated to determine if it played a role in this response. Similar to [Delta]oxyR the [Delta]sigOF mutant was resistant to tBOOH. Interestingly though a double [Delta]sigOF [Delta]oxyR mutant was sensitive to tBOOH in the POST assay and dps expression was reduced as shown by qRTPCR. These results strongly suggest that SigOF is the regulator responsible for dps expression during the POST response.
General note	Presented to the faculty of the Department of Microbiology and Immunology.
General note	Advisor: C. Jeffrey Smith.
General note	Title from PDF t.p. (viewed February 29, 2016).
Dissertation note	Ph.D. East Carolina University 2015.
Bibliography note	Includes bibliographical references.
Technical details	System requirements: Adobe Reader.
Technical details	Mode of access: World Wide Web.

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Role of the iron storage ferritins Dps and DpsL during the prolonged oxidative stress response of Bacteroides fragilis / by Michael I. Betteken.

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