Mapping hisS, the structural gene for histidyl-transfer ribonucleic acid synthetase, in Escherichia coli

Parker, Jack; Fishman, Scott E.

doi:10.1128/jb.138.1.264-267.1979

Cited by 11 publications

(3 citation statements)

References 15 publications

(9 reference statements)

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“…This corresponds to about 0.14 min on the E. coli map (1) or a Pl cotransduction frequency of 0.80 (33). This is very close to the result previously obtained (20). Much of the DNA between glyA and dapE has now been subcloned onto a variety of plasmids, and restriction maps have been made (5,27).…”

Section: Jk98supporting

confidence: 85%

Identification of the purC gene product of Escherichia coli

Parker¹

1984

J Bacteriol

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The purC region of the Escherichia coli chromosome was isolated from in vivo-derived lambda transducing bacteriophages and cloned in high-copy-number plasmids. The product of the purC gene, phosphoribosylaminoimidazolesuccinocarboxamide synthetase, was identified as a protein with an Mr of ca. 27,000. The level of the protein is increased by more than 60-fold in strains carrying the gene on a highcopy-number plasmid. Purine addition represses the enzyme level in both plasmid-and non-plasmidcontaining strains.The de novo biosynthesis of AMP and GMP involves 13 enzymes. In the enteric bacteria the structural genes for 12 of the enzymes have been mapped. Genes purJ, purH, and purD form an operon, as do genes guaB and guaA. The rest of the genes are not tightly linked, although the gene encoding the enzyme catalyzing the third step in purine synthesis has not been located (7).Studies on the regulation of these genes have been slowed by the difficulty of assaying the enzymes and the complexity of metabolic interconversions. It is known that the expression of the pur genes is repressed upon the addition of purines, with hypoxanthine and guanine being corepressors (9). Results by Benson and Gots (2) and Gots et al. (7) indicate that the pur genes, but not the gua operon, may be controlled by a common aporepressor, the product of the purR gene. A protein which binds to different pur gene promoter regions in the presence of either ATP or GTP has been isolated (11). However, the exact nature of the purR gene and its product is still unknown.Identification of the purine biosynthetic enzymes, cloning of structural genes, and formation of gene fusions (13, 31) promise to be of help in the elucidation of regulatory mechanisms. In this paper I describe the isolation and cloning of purC, the structural gene for phosphoribosylaminoimidazolesuccinocarboxamide synthetase (EC 6.3.2.6; PurC), and the identification of this protein on two-dimensional polyacrylamide gels. MATERIALS AND METHODSBacterial, viral, and plasmid strains. The bacterial strains used in this work are described in Table 1. Lysogens of strain JF298 containing X c1857 Sam7 were prepared by Poul J0rgensen. The plasmid cloning vectors used were pBR322 (3) and pBR328 (26). The recombinant plasmids used, pSIU103, pSIU104, pSIU107, and pSIU111, are described below.Media and growth conditions. The complex medium used was L-broth (12) supplemented with 0.2% (wt/vol) glucose or 0.2% maltose when growing cells as recipients for X transduction. The defined medium was the MOPS medium of Neidhardt et al. (17) supplemented with 0.4% glucose, 10 ,ug of thiamine per ml, and, when necessary, 50 ,ug of L-amino acids and 20 ,ug of purine bases per ml. Cultures were grown aerobically in Erlenmeyer flasks with rotary shaking. Growth was monitored with a Zeiss spectrophotometer.Isolation of transducing phage. X d purC transducing phages were isolated by the general procedure described by 712 Schrenk and Weisberg (24) from attA deletion strains lysogenic for A c1857 Sam7. A lysate pr...

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confidence: 85%

Identification of the purC gene product of Escherichia coli

Parker¹

1984

J Bacteriol

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“…One of the genes, purB, encodes a bifunctional enzyme known to catalyze two reactions of the pathway (2). In E. coli, analogous positions have been found for nine of the genes, and the positions for purI and purG are reversed (7,11). purJ mutants have not been identified (1, 9).…”

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Identification of the enzymatic reactions encoded by the purG and purI genes of Escherichia coli

et al. 1983

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The chromosomal locations of the genes purG and purI on the Escherichia coli linkage map are the opposites of those of Salmonella typhimurium. By methods which permit the identification of lesions in any of the five early enzymes of the purine de novo pathway, the gene-enzyme relationships of the purG and purI loci have been reevaluated in these two organisms. The results demonstrate that the relative locations of the genes encoding the two enzymes (phosphoribosylformylglycinamidine synthetase and phosphoribosylaminoimidazole synthetase) are similar in the two organisms. The gene products have been correctly determined in S. typhimurium. The gene products currently listed for the loci in E. coli are incorrect. The E. coli purG locus is equivalent to the S. typhimurium purI locus, and the E. coli purI locus is equivalent to the S. typhimurium purG locus.

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“…The genetic loci which code for the enzymes of the purine pathway are scattered around the E. coli K-12 chromosome, with the exception of a three-gene operon which codes for the second (purD) and the last two genes (purH, purJ) of the IMP pathway (1). A cluster of purine genes, including those which comprise the two-gene operon for GMP synthesis ( guaAB), exist in the 53to 55-min interval on the recalibrated map (1,19,25). The purF gene, the subject of this paper, maps at 49.5 min and is cotransducible with aroC, which lies to the left at 50 min (1).…”

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purF-lac fusion and direction of purF transcription in Escherichia coli

Smith¹,

Gots²

1980

J Bacteriol

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The purF locus codes for the first enzyme, glutamine phosphoribosylpyrophosphate amidotransferase, of the purine biosynthetic pathway. A strain of Escherichia coli K-12 was isolated in which the lac structural genes were fused to the control region of the purF locus. This purF-lac fusion was shown to respond to purine-specific regulatory signals. A plaque-forming lambda transducing phage bearing this purF-lac fusion was isolated. This phage was used to genetically determine the direction of transcription for the pufF locus by two independent means. Results from both methods agreed that the direction of transcription of the purF locus was clockwise on the standard Escherichia coli K-12 genetic map.

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Mapping hisS, the structural gene for histidyl-transfer ribonucleic acid synthetase, in Escherichia coli

Abstract: The structural gene for histidyl-tRNA synthetase was localized to 53.8 min on the Escherichia coli genome. The gene order in this region was determined to be dapE-purC-upp-purG-(guaA, guaB)-hisS-glyA.

Cited by 11 publications

References 15 publications

Identification of the purC gene product of Escherichia coli

Identification of the purC gene product of Escherichia coli

Identification of the enzymatic reactions encoded by the purG and purI genes of Escherichia coli

purF-lac fusion and direction of purF transcription in Escherichia coli

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