{"id":{"repo_id":"wfu","oai_identifier":"oai:wakespace.lib.wfu.edu:10339/14656"},"canonical_url":"https://search.dev.ndltd.org/etd/wfu/oai:wakespace.lib.wfu.edu:10339/14656","repository":{"repo_id":"wfu","name":"Wake Forest University","base_url":"https://wakespace.lib.wfu.edu/oai/request"},"display":{"title":"INVESTIGATION OF UNIQUE COENZYME A BIOSYNTHETIC AND REDOX FUNCTIONS IN Bacillus anthracis","abstract":"Coenzyme A (CoASH) replaces the common tripeptide thiol glutathione as the major low molecular-weight thiol in the human pathogen Bacillus anthracis. A novel type III pantothenate kinase (PanK) catalyzes the first committed step in the CoASH biosynthetic pathway in B. anthracis. The 2.0 Å crystal structure of the B. anthracis PanK (BaPanK) demonstrates that it is a new member of the Acetate and Sugar Kinase/Hsc70/Actin (ASKHA) superfamily and modeling the pantothenate and ATP substrates into the active-site cleft provides a clear rationale for the absence of CoASH feedback inhibition. In addition, bioinformatics analyses indicate a widespread distribution of type III PanK isoforms. The gene encoding BaPanK, coaX, is transcribed as part of a tricistronic operon including hslO and cysK-1 loci, which encode the redox-regulated heat shock protein Hsp33 and cysteine synthase A, respectively. A conditional coaX mutant, in the absence of inducer, demonstrates exponential growth after a lag period of 8 h; this unanticipated result is due to a guanine to adenine suppressor mutation identified in the lac operator. Transcription of the tricistronic coaX-hslO-cysK-1 mRNA is observed in the suppressor mutant, as is observed with wild-type B. anthracis. Therefore, these data support BaPanK as an essential enzyme, thus contributing to its validation as a new antimicrobial target. The intracellular cellular thiol-disulfide redox status of B. anthracis is maintained by coenzyme A-disulfide reductase (BACoADR), which catalyzes the NAD(P)H-dependent reduction of coenzyme A-disulfide (CoAD) to 2 CoASH. B. anthracis also contains an additional CoADR isoform, coenzyme A-disulfide reductase-rhodanese homology domain (CoADR-RHD), which does not catalyze the reduction of CoAD and is implicated in to function in sulfur metabolism. In order to test the physiological contributions of BACoADR and CoADR-RHD in thiol-disulfide redox homeostasis, as well as germination and outgrowth of the B. anthracis endospore, in-frame deletion mutants of their respective encoding genes, cdr and cdrX, were constructed, singly and in combination. Both BACoADR and CoADR-RHD appear to have a role in the morphological transition from the endospore to the vegetative cell. However, CoADR-RHD appears to be the primary enzyme responsible for protection against diamide-induced disulfide stress. CoADRs represent unique members of the pyridine nucleotide disulfide oxidoreductase (PNDOR) family due to a single active-site cysteine, present as a stable Cys-SSCoA mixed disulfide. In order to distinguish CoADRs from the closely related NADH (per)oxidases and enable more accurate identification of CoADR proteins among its larger superfamily, hidden Markov model-based bioinformatics analyses have led to the development of CoADR-specific functional motifs.","abstract_html":"Coenzyme A (CoASH) replaces the common tripeptide thiol glutathione as the major low molecular-weight thiol in the human pathogen Bacillus anthracis. A novel type III pantothenate kinase (PanK) catalyzes the first committed step in the CoASH biosynthetic pathway in B. anthracis. The 2.0 Å crystal structure of the B. anthracis PanK (BaPanK) demonstrates that it is a new member of the Acetate and Sugar Kinase/Hsc70/Actin (ASKHA) superfamily and modeling the pantothenate and ATP substrates into the active-site cleft provides a clear rationale for the absence of CoASH feedback inhibition. In addition, bioinformatics analyses indicate a widespread distribution of type III PanK isoforms. The gene encoding BaPanK, coaX, is transcribed as part of a tricistronic operon including hslO and cysK-1 loci, which encode the redox-regulated heat shock protein Hsp33 and cysteine synthase A, respectively. A conditional coaX mutant, in the absence of inducer, demonstrates exponential growth after a lag period of 8 h; this unanticipated result is due to a guanine to adenine suppressor mutation identified in the lac operator. Transcription of the tricistronic coaX-hslO-cysK-1 mRNA is observed in the suppressor mutant, as is observed with wild-type B. anthracis. Therefore, these data support BaPanK as an essential enzyme, thus contributing to its validation as a new antimicrobial target. The intracellular cellular thiol-disulfide redox status of B. anthracis is maintained by coenzyme A-disulfide reductase (BACoADR), which catalyzes the NAD(P)H-dependent reduction of coenzyme A-disulfide (CoAD) to 2 CoASH. B. anthracis also contains an additional CoADR isoform, coenzyme A-disulfide reductase-rhodanese homology domain (CoADR-RHD), which does not catalyze the reduction of CoAD and is implicated in to function in sulfur metabolism. In order to test the physiological contributions of BACoADR and CoADR-RHD in thiol-disulfide redox homeostasis, as well as germination and outgrowth of the B. anthracis endospore, in-frame deletion mutants of their respective encoding genes, cdr and cdrX, were constructed, singly and in combination. Both BACoADR and CoADR-RHD appear to have a role in the morphological transition from the endospore to the vegetative cell. However, CoADR-RHD appears to be the primary enzyme responsible for protection against diamide-induced disulfide stress. CoADRs represent unique members of the pyridine nucleotide disulfide oxidoreductase (PNDOR) family due to a single active-site cysteine, present as a stable Cys-SSCoA mixed disulfide. In order to distinguish CoADRs from the closely related NADH (per)oxidases and enable more accurate identification of CoADR proteins among its larger superfamily, hidden Markov model-based bioinformatics analyses have led to the development of CoADR-specific functional motifs.","abstract_has_math":false,"creators":["Paige, Carleitta"],"institution":"Wake Forest University","degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2009,"date_issued":"2009-03-19T14:17:45Z","date_published":"2009-03-19T14:17:45Z","updated_at":"2026-07-27T22:00:55Z","subjects":["Bacillus anthracis"],"languages":["en_US"],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/10339/14656","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["Paige, Carleitta"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2009-03-19T14:17:45Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2009-03-19T14:17:45Z"]},{"key":"dc:date.issued","label":"Date","values":["2009-03-19T14:17:45Z"]},{"key":"dc:publisher","label":"Institution","values":["Wake Forest University"]},{"key":"dc:type","label":"Dc Type","values":["Dissertation"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Bacillus anthracis"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en_US"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/10339/14656"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Coenzyme A (CoASH) replaces the common tripeptide thiol glutathione as the major low molecular-weight thiol in the human pathogen Bacillus anthracis. A novel type III pantothenate kinase (PanK) catalyzes the first committed step in the CoASH biosynthetic pathway in B. anthracis. The 2.0 Å crystal structure of the B. anthracis PanK (BaPanK) demonstrates that it is a new member of the Acetate and Sugar Kinase/Hsc70/Actin (ASKHA) superfamily and modeling the pantothenate and ATP substrates into the active-site cleft provides a clear rationale for the absence of CoASH feedback inhibition. In addition, bioinformatics analyses indicate a widespread distribution of type III PanK isoforms. The gene encoding BaPanK, coaX, is transcribed as part of a tricistronic operon including hslO and cysK-1 loci, which encode the redox-regulated heat shock protein Hsp33 and cysteine synthase A, respectively. A conditional coaX mutant, in the absence of inducer, demonstrates exponential growth after a lag period of 8 h; this unanticipated result is due to a guanine to adenine suppressor mutation identified in the lac operator. Transcription of the tricistronic coaX-hslO-cysK-1 mRNA is observed in the suppressor mutant, as is observed with wild-type B. anthracis. Therefore, these data support BaPanK as an essential enzyme, thus contributing to its validation as a new antimicrobial target. The intracellular cellular thiol-disulfide redox status of B. anthracis is maintained by coenzyme A-disulfide reductase (BACoADR), which catalyzes the NAD(P)H-dependent reduction of coenzyme A-disulfide (CoAD) to 2 CoASH. B. anthracis also contains an additional CoADR isoform, coenzyme A-disulfide reductase-rhodanese homology domain (CoADR-RHD), which does not catalyze the reduction of CoAD and is implicated in to function in sulfur metabolism. In order to test the physiological contributions of BACoADR and CoADR-RHD in thiol-disulfide redox homeostasis, as well as germination and outgrowth of the B. anthracis endospore, in-frame deletion mutants of their respective encoding genes, cdr and cdrX, were constructed, singly and in combination. Both BACoADR and CoADR-RHD appear to have a role in the morphological transition from the endospore to the vegetative cell. However, CoADR-RHD appears to be the primary enzyme responsible for protection against diamide-induced disulfide stress. CoADRs represent unique members of the pyridine nucleotide disulfide oxidoreductase (PNDOR) family due to a single active-site cysteine, present as a stable Cys-SSCoA mixed disulfide. In order to distinguish CoADRs from the closely related NADH (per)oxidases and enable more accurate identification of CoADR proteins among its larger superfamily, hidden Markov model-based bioinformatics analyses have led to the development of CoADR-specific functional motifs."]},{"key":"dc:title","label":"Title","values":["INVESTIGATION OF UNIQUE COENZYME A BIOSYNTHETIC AND REDOX FUNCTIONS IN Bacillus anthracis"]}]}],"canonical_facts":{"dc:creator":["Paige, Carleitta"],"dc:date.accessioned":["2009-03-19T14:17:45Z"],"dc:date.available":["2009-03-19T14:17:45Z"],"dc:date.issued":["2009-03-19T14:17:45Z"],"dc:description.abstract":["Coenzyme A (CoASH) replaces the common tripeptide thiol glutathione as the major low molecular-weight thiol in the human pathogen Bacillus anthracis. A novel type III pantothenate kinase (PanK) catalyzes the first committed step in the CoASH biosynthetic pathway in B. anthracis. The 2.0 Å crystal structure of the B. anthracis PanK (BaPanK) demonstrates that it is a new member of the Acetate and Sugar Kinase/Hsc70/Actin (ASKHA) superfamily and modeling the pantothenate and ATP substrates into the active-site cleft provides a clear rationale for the absence of CoASH feedback inhibition. In addition, bioinformatics analyses indicate a widespread distribution of type III PanK isoforms. The gene encoding BaPanK, coaX, is transcribed as part of a tricistronic operon including hslO and cysK-1 loci, which encode the redox-regulated heat shock protein Hsp33 and cysteine synthase A, respectively. A conditional coaX mutant, in the absence of inducer, demonstrates exponential growth after a lag period of 8 h; this unanticipated result is due to a guanine to adenine suppressor mutation identified in the lac operator. Transcription of the tricistronic coaX-hslO-cysK-1 mRNA is observed in the suppressor mutant, as is observed with wild-type B. anthracis. Therefore, these data support BaPanK as an essential enzyme, thus contributing to its validation as a new antimicrobial target. The intracellular cellular thiol-disulfide redox status of B. anthracis is maintained by coenzyme A-disulfide reductase (BACoADR), which catalyzes the NAD(P)H-dependent reduction of coenzyme A-disulfide (CoAD) to 2 CoASH. B. anthracis also contains an additional CoADR isoform, coenzyme A-disulfide reductase-rhodanese homology domain (CoADR-RHD), which does not catalyze the reduction of CoAD and is implicated in to function in sulfur metabolism. In order to test the physiological contributions of BACoADR and CoADR-RHD in thiol-disulfide redox homeostasis, as well as germination and outgrowth of the B. anthracis endospore, in-frame deletion mutants of their respective encoding genes, cdr and cdrX, were constructed, singly and in combination. Both BACoADR and CoADR-RHD appear to have a role in the morphological transition from the endospore to the vegetative cell. However, CoADR-RHD appears to be the primary enzyme responsible for protection against diamide-induced disulfide stress. CoADRs represent unique members of the pyridine nucleotide disulfide oxidoreductase (PNDOR) family due to a single active-site cysteine, present as a stable Cys-SSCoA mixed disulfide. In order to distinguish CoADRs from the closely related NADH (per)oxidases and enable more accurate identification of CoADR proteins among its larger superfamily, hidden Markov model-based bioinformatics analyses have led to the development of CoADR-specific functional motifs."],"dc:identifier.uri":["http://hdl.handle.net/10339/14656"],"dc:language.iso":["en_US"],"dc:publisher":["Wake Forest University"],"dc:subject":["Bacillus anthracis"],"dc:title":["INVESTIGATION OF UNIQUE COENZYME A BIOSYNTHETIC AND REDOX FUNCTIONS IN Bacillus anthracis"],"dc:type":["Dissertation"]},"updated_at":"2026-07-27T22:00:55Z"}