{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/20057"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/20057","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Autogenous transcriptional attenuation of de novo pyrimidine biosynthesis in Bacillus subtilis","abstract":"Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:41:16Z Item is restricted indefinitely.","abstract_html":"Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:41:16Z Item is restricted indefinitely.","abstract_has_math":false,"creators":["Turner, Robert Joseph"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Biochemistry","degree_department":null,"school":null,"contributors":["Switzer, Robert L."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-05-07T12:27:27Z","date_published":"2011-05-07T12:27:27Z","updated_at":"2026-07-22T22:25:15Z","subjects":["Biology, Molecular","Biology, Microbiology","Chemistry, Biochemistry"],"languages":["eng"],"rights":["Copyright 1996 Turner, Robert Joseph"],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["9780591089684","AAI9702694","(UMI)AAI9702694"],"render_values":[{"text":"9780591089684","href":null,"code":true},{"text":"AAI9702694","href":null,"code":true},{"text":"(UMI)AAI9702694","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/20057","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Switzer, Robert L."]},{"key":"dc:creator","label":"Author","values":["Turner, Robert Joseph"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011-05-07T12:27:27Z","10000-01-01","1996"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biochemistry"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Biology, Molecular","Biology, Microbiology","Chemistry, Biochemistry"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 1996 Turner, Robert Joseph"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["9780591089684","AAI9702694","(UMI)AAI9702694","http://hdl.handle.net/2142/20057"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:41:16Z Item is restricted indefinitely.","The genes encoding the enzymes for de novo pyrimidine biosynthesis in the gram-positive microorganism Bacillus subtilis were previously shown to be clustered at 139 min on its chromosome, transcribed from a single promoter and regulated at the level of transcription by the availability of pyrimidines.","A complete transcript of the B. subtilis pyr operon contains the following elements in 5$\\sp\\prime$ to 3$\\sp\\prime$ order: a 151 nt untranslated leader; pyrR, encoding a 20 kDa protein; a 173 nt intercistronic region; pyrP, encoding a 46 kDa protein; a 145 nt intercistronic region; followed by 8 overlapping cistrons encoding all of the 6 enzymes for de novo pyrimidine biosynthesis. Transcription is controlled by the availability of pyrimidines via an attenuation mechanism. Computer analysis of potential RNA structures revealed that there are three rho-independent transcription terminators within the operon, each of which is preceded by another stem-loop structure, the antiterminator, whose formation would prevent formation of the terminator stem-loop. These are located in the leader, pyrR-pyrP intercistronic, and pyrP-pyrB intercistronic regions. Each antiterminator contains a 50 base conserved sequence in its promoter proximal half. Various transcriptional fusions of the pyr promoter and surrounding sequences to promoterless reporter genes support an attenuation mechanism whereby when pyrimidines are abundant, the PyrR protein binds to the conserved sequence in the pyr mRNA and disrupts the antiterminator, permitting terminator hairpin formation and promoting transcription termination.","The regulatory activity of PyrR was confirmed when deletion of pyrR from the chromosome resulted in the constitutive elevated expression of aspartate transcarbamylase, which is encoded by pyrB, the third gene in the operon. Complementation of an E. coli upp mutant, as well as direct enzymatic assay, has demonstrated that pyrR also confers uracil phosphoribosyltransferase (UPRTase) activity. Analysis of pyrR and upp deletion mutants demonstrated that upp, not pyrR, encodes the quantitatively important UPRTase activity. Complementation of B. subtilis furA2, a mutant defective in uracil transport, and amino acid sequence alignment with the E. coli uracil permease strongly suggests pyrP encodes an integral membrane uracil permease.","This work has made it possible to biochemically characterize the regulation of the B. subtilis pyr operon. The availability of purified PyrR permits the analysis of the RNA-binding and UPRTase properties of PyrR, the reconstruction of B. subtilis pyr attenuation in vitro, and the determination of the high resolution structure of PyrR. (Abstract shortened by UMI.)","Made available in DSpace on 2011-05-07T12:27:27Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9702694.pdf: 9326987 bytes, checksum: 3ae8697e219f06e2d08632fd501f2021 (MD5) Previous issue date: 1996","Restriction data tranferred 2014-07-01T11:17:49-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: ETDs are only available to UIUC Users without author permission","ETDs are only available to UIUC Users without author permission","U of I Only"]},{"key":"dc:title","label":"Title","values":["Autogenous transcriptional attenuation of de novo pyrimidine biosynthesis in Bacillus subtilis"]}]}],"canonical_facts":{"dc:contributor":["Switzer, Robert L."],"dc:creator":["Turner, Robert Joseph"],"dc:date":["2011-05-07T12:27:27Z","10000-01-01","1996"],"dc:description":["Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:41:16Z Item is restricted indefinitely.","The genes encoding the enzymes for de novo pyrimidine biosynthesis in the gram-positive microorganism Bacillus subtilis were previously shown to be clustered at 139 min on its chromosome, transcribed from a single promoter and regulated at the level of transcription by the availability of pyrimidines.","A complete transcript of the B. subtilis pyr operon contains the following elements in 5$\\sp\\prime$ to 3$\\sp\\prime$ order: a 151 nt untranslated leader; pyrR, encoding a 20 kDa protein; a 173 nt intercistronic region; pyrP, encoding a 46 kDa protein; a 145 nt intercistronic region; followed by 8 overlapping cistrons encoding all of the 6 enzymes for de novo pyrimidine biosynthesis. Transcription is controlled by the availability of pyrimidines via an attenuation mechanism. Computer analysis of potential RNA structures revealed that there are three rho-independent transcription terminators within the operon, each of which is preceded by another stem-loop structure, the antiterminator, whose formation would prevent formation of the terminator stem-loop. These are located in the leader, pyrR-pyrP intercistronic, and pyrP-pyrB intercistronic regions. Each antiterminator contains a 50 base conserved sequence in its promoter proximal half. Various transcriptional fusions of the pyr promoter and surrounding sequences to promoterless reporter genes support an attenuation mechanism whereby when pyrimidines are abundant, the PyrR protein binds to the conserved sequence in the pyr mRNA and disrupts the antiterminator, permitting terminator hairpin formation and promoting transcription termination.","The regulatory activity of PyrR was confirmed when deletion of pyrR from the chromosome resulted in the constitutive elevated expression of aspartate transcarbamylase, which is encoded by pyrB, the third gene in the operon. Complementation of an E. coli upp mutant, as well as direct enzymatic assay, has demonstrated that pyrR also confers uracil phosphoribosyltransferase (UPRTase) activity. Analysis of pyrR and upp deletion mutants demonstrated that upp, not pyrR, encodes the quantitatively important UPRTase activity. Complementation of B. subtilis furA2, a mutant defective in uracil transport, and amino acid sequence alignment with the E. coli uracil permease strongly suggests pyrP encodes an integral membrane uracil permease.","This work has made it possible to biochemically characterize the regulation of the B. subtilis pyr operon. The availability of purified PyrR permits the analysis of the RNA-binding and UPRTase properties of PyrR, the reconstruction of B. subtilis pyr attenuation in vitro, and the determination of the high resolution structure of PyrR. (Abstract shortened by UMI.)","Made available in DSpace on 2011-05-07T12:27:27Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9702694.pdf: 9326987 bytes, checksum: 3ae8697e219f06e2d08632fd501f2021 (MD5) Previous issue date: 1996","Restriction data tranferred 2014-07-01T11:17:49-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: ETDs are only available to UIUC Users without author permission","ETDs are only available to UIUC Users without author permission","U of I Only"],"dc:identifier":["9780591089684","AAI9702694","(UMI)AAI9702694","http://hdl.handle.net/2142/20057"],"dc:language":["eng"],"dc:rights":["Copyright 1996 Turner, Robert Joseph"],"dc:subject":["Biology, Molecular","Biology, Microbiology","Chemistry, Biochemistry"],"dc:title":["Autogenous transcriptional attenuation of de novo pyrimidine biosynthesis in Bacillus subtilis"],"dc:type":["text"],"thesis:degree_discipline":["Biochemistry"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:25:15Z"}