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Peggy J. Farnham - One of the best experts on this subject based on the ideXlab platform.
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Start site selection at the TATA-less Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase promoter.
The Journal of biological chemistry, 1994Co-Authors: Richard Kollmar, Kristine A. Sukow, Stephen K. Sponagle, Peggy J. FarnhamAbstract:Abstract Transcription of the Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase (CAD) gene from the Syrian hamster, Mesocricetus auratus, starts at a single major site. We characterized the cis-acting elements that position RNA polymerase II at the correct start site in the CAD promoter. Sequence alignment showed that the CAD promoter lacks a TATA box, but contains a consensus initiator. Mutational analysis of the CAD promoter demonstrated that the sequences between -81 and +26 were sufficient for accurate and efficient transcription in vitro and in vivo; binding sites for the transcription factor Sp1 around -70 and -49 were necessary for transcriptional activity. The binding site at -49 directed initiation about 50 base pairs downstream. A ubiquitous activator protein, Honk, bound to the CAD promoter between -30 and -12, but did not participate in start site selection. The sequences around +1, which contain the consensus initiator, contributed to promoter activity; however, the presence of a consensus initiator in this region was neither necessary nor sufficient for transcription. We concluded from these results that the Sp1 binding site at -49 substituted for the missing TATA box and played a major role in start site selection at the CAD promoter.
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start site selection at the tata less Carbamoyl Phosphate Synthase glutamine hydrolyzing aspartate Carbamoyltransferase dihydroorotase promoter
Journal of Biological Chemistry, 1994Co-Authors: Richard Kollmar, Kristine A. Sukow, Stephen K. Sponagle, Peggy J. FarnhamAbstract:Abstract Transcription of the Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase (CAD) gene from the Syrian hamster, Mesocricetus auratus, starts at a single major site. We characterized the cis-acting elements that position RNA polymerase II at the correct start site in the CAD promoter. Sequence alignment showed that the CAD promoter lacks a TATA box, but contains a consensus initiator. Mutational analysis of the CAD promoter demonstrated that the sequences between -81 and +26 were sufficient for accurate and efficient transcription in vitro and in vivo; binding sites for the transcription factor Sp1 around -70 and -49 were necessary for transcriptional activity. The binding site at -49 directed initiation about 50 base pairs downstream. A ubiquitous activator protein, Honk, bound to the CAD promoter between -30 and -12, but did not participate in start site selection. The sequences around +1, which contain the consensus initiator, contributed to promoter activity; however, the presence of a consensus initiator in this region was neither necessary nor sufficient for transcription. We concluded from these results that the Sp1 binding site at -49 substituted for the missing TATA box and played a major role in start site selection at the CAD promoter.
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Heat sensitivity and Sp1 activation of complex formation at the Syrian hamster Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase promoter in vitro.
The Journal of biological chemistry, 1992Co-Authors: Richard Kollmar, Mary J. Lindstrom, Peggy J. FarnhamAbstract:Abstract To study the regulation of transcription of the Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase (CAD) gene from the Syrian hamster, Mesocricetus auratus, we developed a homologous in vitro transcription system on the basis of nuclear extract from Syrian hamster kidney cells. We optimized the reaction temperature and the concentrations of DNA template, KCl, and MgCl2 simultaneously with the response surface method and found an unusually low temperature optimum of 20 degrees C. We therefore investigated whether CAD transcription in vitro depended on a heat-labile component of nuclear extract. Preincubating extract alone at 30 degrees C reduced transcription from the CAD promoter but not from the major late promoter of adenovirus 2. The formation of stable initiation complexes at the CAD promoter was diminished in heat-treated extract; run-off transcripts, however, accumulated at the same rate as in untreated extract. The heat sensitivity of complex formation correlated with the heat sensitivity of DNA binding by transcription factor Sp1, which binds to two sites in the CAD promoter; moreover, both preformed initiation complexes and DNA-bound Sp1 were heat-resistant. Adding purified Sp1 to heat-treated extract restored complex formation. We propose that Sp1 activates CAD transcription by stabilizing initiation complexes at the CAD promoter.
Richard Kollmar - One of the best experts on this subject based on the ideXlab platform.
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Start site selection at the TATA-less Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase promoter.
The Journal of biological chemistry, 1994Co-Authors: Richard Kollmar, Kristine A. Sukow, Stephen K. Sponagle, Peggy J. FarnhamAbstract:Abstract Transcription of the Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase (CAD) gene from the Syrian hamster, Mesocricetus auratus, starts at a single major site. We characterized the cis-acting elements that position RNA polymerase II at the correct start site in the CAD promoter. Sequence alignment showed that the CAD promoter lacks a TATA box, but contains a consensus initiator. Mutational analysis of the CAD promoter demonstrated that the sequences between -81 and +26 were sufficient for accurate and efficient transcription in vitro and in vivo; binding sites for the transcription factor Sp1 around -70 and -49 were necessary for transcriptional activity. The binding site at -49 directed initiation about 50 base pairs downstream. A ubiquitous activator protein, Honk, bound to the CAD promoter between -30 and -12, but did not participate in start site selection. The sequences around +1, which contain the consensus initiator, contributed to promoter activity; however, the presence of a consensus initiator in this region was neither necessary nor sufficient for transcription. We concluded from these results that the Sp1 binding site at -49 substituted for the missing TATA box and played a major role in start site selection at the CAD promoter.
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start site selection at the tata less Carbamoyl Phosphate Synthase glutamine hydrolyzing aspartate Carbamoyltransferase dihydroorotase promoter
Journal of Biological Chemistry, 1994Co-Authors: Richard Kollmar, Kristine A. Sukow, Stephen K. Sponagle, Peggy J. FarnhamAbstract:Abstract Transcription of the Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase (CAD) gene from the Syrian hamster, Mesocricetus auratus, starts at a single major site. We characterized the cis-acting elements that position RNA polymerase II at the correct start site in the CAD promoter. Sequence alignment showed that the CAD promoter lacks a TATA box, but contains a consensus initiator. Mutational analysis of the CAD promoter demonstrated that the sequences between -81 and +26 were sufficient for accurate and efficient transcription in vitro and in vivo; binding sites for the transcription factor Sp1 around -70 and -49 were necessary for transcriptional activity. The binding site at -49 directed initiation about 50 base pairs downstream. A ubiquitous activator protein, Honk, bound to the CAD promoter between -30 and -12, but did not participate in start site selection. The sequences around +1, which contain the consensus initiator, contributed to promoter activity; however, the presence of a consensus initiator in this region was neither necessary nor sufficient for transcription. We concluded from these results that the Sp1 binding site at -49 substituted for the missing TATA box and played a major role in start site selection at the CAD promoter.
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Heat sensitivity and Sp1 activation of complex formation at the Syrian hamster Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase promoter in vitro.
The Journal of biological chemistry, 1992Co-Authors: Richard Kollmar, Mary J. Lindstrom, Peggy J. FarnhamAbstract:Abstract To study the regulation of transcription of the Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase (CAD) gene from the Syrian hamster, Mesocricetus auratus, we developed a homologous in vitro transcription system on the basis of nuclear extract from Syrian hamster kidney cells. We optimized the reaction temperature and the concentrations of DNA template, KCl, and MgCl2 simultaneously with the response surface method and found an unusually low temperature optimum of 20 degrees C. We therefore investigated whether CAD transcription in vitro depended on a heat-labile component of nuclear extract. Preincubating extract alone at 30 degrees C reduced transcription from the CAD promoter but not from the major late promoter of adenovirus 2. The formation of stable initiation complexes at the CAD promoter was diminished in heat-treated extract; run-off transcripts, however, accumulated at the same rate as in untreated extract. The heat sensitivity of complex formation correlated with the heat sensitivity of DNA binding by transcription factor Sp1, which binds to two sites in the CAD promoter; moreover, both preformed initiation complexes and DNA-bound Sp1 were heat-resistant. Adding purified Sp1 to heat-treated extract restored complex formation. We propose that Sp1 activates CAD transcription by stabilizing initiation complexes at the CAD promoter.
Huasong Zou - One of the best experts on this subject based on the ideXlab platform.
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Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri
BMC Microbiology, 2015Co-Authors: Tao Zhuo, Jing Guo, Xue Song, Wei Rou, Xiaojing Fan, Gicharu Gibson Kamau, Huasong ZouAbstract:Background The carA and carB genes code the small and large subunits of Carbamoyl-Phosphate Synthase (CPS) that responsible for arginine and pyrimidine production. The purpose of this work was to study the gene organization and expression pattern of carAB operon, and the biological functions of carA and carB genes in Xanthomonas citri subsp. citri.
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The small and large subunits of Carbamoyl-Phosphate Synthase exhibit diverse contributions to pathogenicity in Xanthomonas citri subsp. citri
Journal of Integrative Agriculture, 2015Co-Authors: Jing Guo, Xue Song, Lifang Zou, Huasong Zou, Gongyou ChenAbstract:Abstract Carbamoyl-Phosphate Synthase plays a vital role in the carbon and nitrogen metabolism cycles. In Xanthomonas citri subsp. citri, carA and carB encode the small and large subunits of Carbamoyl-Phosphate Synthase, respectively. The deletion mutation of the coding regions revealed that carA did not affect any of the phenotypes, while carB played multiple roles in pathogenicity. The deletion of carB rendered the loss of pathogenicity in host plants and the ability to induce a hypersensitive reaction in the non-hosts. Quantitative reverse transcription-PCR assays indicated that 11 hrp genes coding the type III secretion system were suppressed when interacting with citrus plants. The mutation in carB also affected bacterial utilization of several carbon and nitrogen resources in minimal medium MMX and extracellular enzyme activities. These data demonstrated that only the large subunit of Carbamoyl-Phosphate Synthase was essential for canker development by X. citri subsp. citri.
Jing Guo - One of the best experts on this subject based on the ideXlab platform.
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Molecular study on the carAB operon reveals that carB gene is required for swimming and biofilm formation in Xanthomonas citri subsp. citri
BMC Microbiology, 2015Co-Authors: Tao Zhuo, Jing Guo, Xue Song, Wei Rou, Xiaojing Fan, Gicharu Gibson Kamau, Huasong ZouAbstract:Background The carA and carB genes code the small and large subunits of Carbamoyl-Phosphate Synthase (CPS) that responsible for arginine and pyrimidine production. The purpose of this work was to study the gene organization and expression pattern of carAB operon, and the biological functions of carA and carB genes in Xanthomonas citri subsp. citri.
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The small and large subunits of Carbamoyl-Phosphate Synthase exhibit diverse contributions to pathogenicity in Xanthomonas citri subsp. citri
Journal of Integrative Agriculture, 2015Co-Authors: Jing Guo, Xue Song, Lifang Zou, Huasong Zou, Gongyou ChenAbstract:Abstract Carbamoyl-Phosphate Synthase plays a vital role in the carbon and nitrogen metabolism cycles. In Xanthomonas citri subsp. citri, carA and carB encode the small and large subunits of Carbamoyl-Phosphate Synthase, respectively. The deletion mutation of the coding regions revealed that carA did not affect any of the phenotypes, while carB played multiple roles in pathogenicity. The deletion of carB rendered the loss of pathogenicity in host plants and the ability to induce a hypersensitive reaction in the non-hosts. Quantitative reverse transcription-PCR assays indicated that 11 hrp genes coding the type III secretion system were suppressed when interacting with citrus plants. The mutation in carB also affected bacterial utilization of several carbon and nitrogen resources in minimal medium MMX and extracellular enzyme activities. These data demonstrated that only the large subunit of Carbamoyl-Phosphate Synthase was essential for canker development by X. citri subsp. citri.
Kristine A. Sukow - One of the best experts on this subject based on the ideXlab platform.
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Start site selection at the TATA-less Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase promoter.
The Journal of biological chemistry, 1994Co-Authors: Richard Kollmar, Kristine A. Sukow, Stephen K. Sponagle, Peggy J. FarnhamAbstract:Abstract Transcription of the Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase (CAD) gene from the Syrian hamster, Mesocricetus auratus, starts at a single major site. We characterized the cis-acting elements that position RNA polymerase II at the correct start site in the CAD promoter. Sequence alignment showed that the CAD promoter lacks a TATA box, but contains a consensus initiator. Mutational analysis of the CAD promoter demonstrated that the sequences between -81 and +26 were sufficient for accurate and efficient transcription in vitro and in vivo; binding sites for the transcription factor Sp1 around -70 and -49 were necessary for transcriptional activity. The binding site at -49 directed initiation about 50 base pairs downstream. A ubiquitous activator protein, Honk, bound to the CAD promoter between -30 and -12, but did not participate in start site selection. The sequences around +1, which contain the consensus initiator, contributed to promoter activity; however, the presence of a consensus initiator in this region was neither necessary nor sufficient for transcription. We concluded from these results that the Sp1 binding site at -49 substituted for the missing TATA box and played a major role in start site selection at the CAD promoter.
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start site selection at the tata less Carbamoyl Phosphate Synthase glutamine hydrolyzing aspartate Carbamoyltransferase dihydroorotase promoter
Journal of Biological Chemistry, 1994Co-Authors: Richard Kollmar, Kristine A. Sukow, Stephen K. Sponagle, Peggy J. FarnhamAbstract:Abstract Transcription of the Carbamoyl-Phosphate Synthase (glutamine-hydrolyzing)/aspartate Carbamoyltransferase/dihydroorotase (CAD) gene from the Syrian hamster, Mesocricetus auratus, starts at a single major site. We characterized the cis-acting elements that position RNA polymerase II at the correct start site in the CAD promoter. Sequence alignment showed that the CAD promoter lacks a TATA box, but contains a consensus initiator. Mutational analysis of the CAD promoter demonstrated that the sequences between -81 and +26 were sufficient for accurate and efficient transcription in vitro and in vivo; binding sites for the transcription factor Sp1 around -70 and -49 were necessary for transcriptional activity. The binding site at -49 directed initiation about 50 base pairs downstream. A ubiquitous activator protein, Honk, bound to the CAD promoter between -30 and -12, but did not participate in start site selection. The sequences around +1, which contain the consensus initiator, contributed to promoter activity; however, the presence of a consensus initiator in this region was neither necessary nor sufficient for transcription. We concluded from these results that the Sp1 binding site at -49 substituted for the missing TATA box and played a major role in start site selection at the CAD promoter.