Cloning, nucleotide sequence and regulation of the Salmonella typhimurium pyrD gene encoding dihydroorotate dehydrogenase.

The Salmonella typhimurium pyrD gene encoding dihydroorotate dehydrogenase was cloned and sequenced. In total, a sequence of 1286 nucleotide pairs was determined wherein a single open-reading-frame of 1011 bp, encoding a polypeptide of 336 amino acids having 95% similarity with the Escherichia coli pyrD gene product, was identified. A region of hyphenated-dyad symmetry exists within the leader region affording the potential for the formation of a stable secondary structure in the 5' end of the transcript. Mutations from several regulatory mutants were located within the region of dyad symmetry which would impart changes in the transcript within the putative secondary structure, implicating the secondary structure in regulation. Primer extension analysis revealed multiple transcriptional start sites located six to nine nucleotides downstream from the Pribnow box, with the primary initiation site differing in repressing and derepressing growth conditions. The results are discussed in terms of a translational attenuation model for regulation of pyrD expression.

[1]  K. Jensen.  Regulation of Salmonella typhimurium pyr gene expression: effect of changing both purine and pyrimidine nucleotide pools. , 1989, Journal of general microbiology.

[2]  R. Rolfes,et al.  Escherichia coli gene purR encoding a repressor protein for purine nucleotide synthesis. Cloning, nucleotide sequence, and interaction with the purF operator. , 1988, The Journal of biological chemistry.

[3]  L. Gold,et al.  Posttranscriptional regulatory mechanisms in Escherichia coli. , 1988, Annual review of biochemistry.

[4]  F. Nargang,et al.  Cloning, nucleotide sequence and expression of the pyrBI operon of Salmonella typhimurium LT2. , 1987, European journal of biochemistry.

[5]  C. Turnbough,et al.  Nucleotide sequence and expression of the pyrC gene of Escherichia coli K-12 , 1987, Journal of bacteriology.

[6]  B. Wanner.  Molecular cloning of Mu d(bla lacZ) transcriptional and translational fusions , 1987, Journal of bacteriology.

[7]  M. Hartley,et al.  A low-copy-number vector utilizing beta-galactosidase for the analysis of gene control elements. , 1987, Gene.

[8]  D. Bäckström,et al.  Nucleotide sequence of the structural gene for dihydroorotase of Escherichia coli K12. , 1986, European journal of biochemistry.

[9]  J. Neuhard,et al.  Cloning and structural characterization of the Salmonella typhimurium pyrC gene encoding dihydroorotase. , 1986, European journal of biochemistry.

[10]  J. Neuhard,et al.  Isolation and characterization of pyrimidine mutants of Salmonella typhimurium altered in expression of pyrC, pyrD, and pyrE. , 1985, Canadian journal of microbiology.

[11]  K. Jensen,et al.  Nucleotide sequence of the pyrD gene of Escherichia coli and characterization of the flavoprotein dihydroorotate dehydrogenase. , 1985, European journal of biochemistry.

[12]  J. Neuhard,et al.  Cloning and characterization of the pyrE gene and of PyrE::Mud1 (Ap lac) fusions from Salmonella typhimurium. , 1985, European journal of biochemistry.

[13]  W. Fiers,et al.  Alterations upstream from the Shine-Dalgarno region and their effect on bacterial gene expression. , 1985, Gene.

[14]  R. Dale,et al.  A rapid single-stranded cloning strategy for producing a sequential series of overlapping clones for use in DNA sequencing: application to sequencing the corn mitochondrial 18 S rDNA. , 1985, Plasmid.

[15]  N. Minton,et al.  Improved plasmid vectors for the isolation of translational lac gene fusions. , 1984, Gene.

[16]  K. Jensen,et al.  Structure of the Escherichia coli pyrE operon and control of pyrE expression by a UTP modulated intercistronic attentuation. , 1984, The EMBO journal.

[17]  D. Mount,et al.  Rapid mutational analysis of regulatory loci in Escherichia coli K-12 using bacteriophage M13. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[18]  J. Davison,et al.  Vectors with restriction-site banks. I. pJRD158, a 3903-bp plasmid containing 28 unique cloning sites. , 1984, Gene.

[19]  K. Jensen,et al.  Studies on the structure and expression of Escherichia coli pyrC, pyrD, and pyrF using the cloned genes. , 1984, European journal of biochemistry.

[20]  J. Messing.  [2] New M13 vectors for cloning , 1983 .

[21]  C. Turnbough,et al.  Attenuation control of pyrBI operon expression in Escherichia coli K-12. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Michael Zuker,et al.  Optimal computer folding of large RNA sequences using thermodynamics and auxiliary information , 1981, Nucleic Acids Res..

[23]  K. Jensen.  Apparent involvement of purines in the control of expression of Salmonella typhimurium pyr genes: analysis of a leaky guaB mutant resistant to pyrimidine analogs , 1979, Journal of bacteriology.

[24]  D. Karibian.  Dihydroorotate dehydrogenase (Escherichia coli). , 1978, Methods in enzymology.

[25]  R. A. Kelln,et al.  Pyrimidine biosynthetic enzymes of Salmonella typhimurium, repressed specifically by growth in the presence of cytidine , 1975, Journal of bacteriology.

[26]  M. Schwartz,et al.  Control of expression of the pyr genes in Salmonella typhimurium: effects of variations in uridine and cytidine nucleotide pools , 1975, Journal of bacteriology.

[27]  Jeffrey H. Miller.  Experiments in molecular genetics , 1972 .

[28]  M. L. Taylor,et al.  ENZYMES OF THE PYRIMIDINE PATHWAY IN ESCHERICHIA COLI II , 1964, Journal of bacteriology.