Browse > Article

Identification of Potential Corynebacterium ammoniagenes Purine Gene Regulators Using the pur-lacZ Reporter in Escherichia coli  

HAN , RI-NA (Department of Biotechnology, Yonsei University)
CHO, ICK-HYUN (Department of Biotechnology, Yonsei University)
CHUNG, SUNG-OH (R&D Center for Bioproducts, CJ Co)
HAN, JONG-KWON (R&D Center for Bioproducts, CJ Co)
LEE, JIN-HOO (R&D Center for Bioproducts, CJ Co)
KIM, SOO-KI (Department of Animal Sciences and Environment, Konkuk University)
CHOI, KANG-YELL (Department of Biotechnology, Yonsei University)
Publication Information
Journal of Microbiology and Biotechnology / v.14, no.6, 2004 , pp. 1249-1255 More about this Journal
Abstract
This study has developed Corynebacterium ammoniagenes (c. ammoniagenes) purine gene transcriptional reporters (purF-lacZ and purE-lacZ) that function in Escherichia coli (E. coli) DH5a. After transformation of a C. ammoniagenes gDNA library into E. coli cells harboring either purF-lacZ or purE-lacZ, C. ammoniagenes clones were obtained that repress purF-lacZ and purE-lacZ gene expression. The potential purE and purF regulatory genes are homologous to the genes encoding transcription regulators, the regulatory subunit of RNA polymerase, and genes for purine nucleotide biosynthesis of various bacteria. The C. ammoniagenes purE-lacZ and purF-lacZ reporters were repressed by adenine and guanine within E. coli, indicating similarity in the regulatory mechanism of purine biosynthesis in C. ammoniagenes and E. coli. Gene regulation of pur-lacZ by adenine and guanine was partly abolished in cells expressing potential purine regulatory genes, indicating functionality of the purine gene regulators in repression of purE-lacZ and purF-lacZ. The purE-lacZ and purF-lacZ reporters can be used for the screening of genes involved in the regulation of the de novo synthesis of the purine nucleotides.
Keywords
Corynebacterium ammoniagenes; de novo purine nucleotide biosynthesis; reporter system; purE; purF; reporter; repressor;
Citations & Related Records

Times Cited By Web Of Science : 0  (Related Records In Web of Science)
연도 인용수 순위
  • Reference
1 Choi, K. Y. and H. Zalkin. 1990. Regulation of Escherichia coli pyrC by the purine regulon repressor protein. J. Bacteriol. 172: 3201-3207
2 Choi, K. Y., F. Lu, and H. Zalkin. 1994. Mutagenesis of amino acid residues required for binding of corepressors to the purine repressor. J. Biol. Chem. 269: 24066- 24072
3 Chung, S. O., J. H. Lee, S. Y. Lee, and D. S. Lee. 1996. Genomic organization of purK and purE in Brevibacterium ammoniagenes ATCC6872: Pure locus provides a clue for genomic evolution. FEMS Microbiol. Lett. 137: 265-268
4 Ebbole, D. J. and H. Zalkin. 1986. Interaction of a putative repressor protein with an extended control region of the Bacillus subtilis pur operon. J. Biol. Chem. 264: 3553-3561
5 Eikmanns, B. J., E. Klentz, W. Liebl, and H. A. Shm. 1991. A family of Corynebacterium glutamicum/Escherichia coli shuttle vectors for cloning, controlled gene expression, and promoter probing. Gene 2: 93-98
6 Meng, L. M., M. Kilstruop, and P. Nygaard. 1990. Autoregulation of PurR repressor synthesis and involvement of purR in the regulation of purB, purC, purL, purMN, and guaBA expression in Escherichia coli. Eur. J. Biochem. 187: 373-379
7 Mongkolsuk, S., E. J. Duvall, and P. S. Lovett. 1985. Transcription termination signal for the cat-86 indicator gene in a Bacillus subtilis promoter-cloning plasmid. Gene 37: 83-90
8 Nagy, M. 1970. Regulation of the biosynthesis of purine nucleotides in Schizosaccharomyces pombe: I. Properties of the phosphoribosylpyrophosphate: glutamine amidotransferase of the wild strain and of a mutant desensitized towards feedback modifiers. Biochim. Biophys. Acta 198: 471-481
9 Perna, N. T., G. Plunkett III, V. Burland, B. Mau, J. D. Glasner, D. J. Rose, G. F. Mayhew, P. S. Evans, J. Gregor, H. A. Kirkpatrick, G. Posfai, J. Hackett, S. Klink, A. Boutin, Y. Shao, L. Miller, E. J. Grotbeck, N. W. Davis, A. Lim, E. Dimalanta, K. Potamousis, J. Apodaca, T. S. Anantharaman, J. Lin, G. Yen, D. C. Schwartz, R. A. Welch, and F. R. Blattner. 2001. Genome sequence of enterohaemorrhagic Escherichia coli O157:H7. Nature 409: 529-533
10 Saxild, H. H. and P. Nygaard. 1998. Gene-enzyme relationships of the purine biosynthetic pathway in Bacillus subtilis. Mol. Gen. Genet. 211: 160-167
11 Steiert, J. G., R. J. Rofels, H. Zalkin, and G. V. Stauffer. 1990. Regulation of Escherichia coli glyA gene by the purR gene product. J. Bacteriol. 172: 3799-3830
12 Weng, M. and H. Zalkin. 2000. Mutations in the Bacillus subtilis purine repressor that perturb PRPP effector function in vitro and in vivo. Curr. Microbiol. 41: 56-59
13 Zalkin, H. 1999. De novo purine nucleotide synthesis, pp. 335-341. In A. L. Sonenshein, J. A. Hoch, and R. Losick (eds.), Bacillus subtilis and Other Gram-Positive Bacteria: Biochemistry, Physiology, and Molecular Genetics. American Society for Microbiology, Washington, D.C., U.S.A
14 Wilson, H. R. and C. L. Turnbough, Jr. 1990. Role of the purine repressor in the regulation of pyrimidine gene expression in Escherichia coli K-12. J. Bacteriol. 172: 3208-3213
15 Choi, K. Y. and H. Zalkin. 1992. Structural characterization and corepressor binding of Escherichia coli purine repressor. J. Bacteriol. 174: 6207-6214
16 Rappu, P., B. S. Shin, H. Zalkin, and P. Mäntsälä. 1999. A role for a highly conserved protein of unknown function in regulation of Bacillus subtilis purA by the purine repressor. J. Bacteriol. 181: 3810-3815
17 Got, J. S. and C. S. Gollub. 1959. Purine analogs as feedback inhibitors. Proc. Soc. Exp. Biol. Med. 101: 641-643
18 Patek, M., B. J. Eikmanns, J. Patek, and H. Sahm. 1996. Promoters from Corynebacterium glutamicum: Cloning, molecular analysis and search for a consensus motif. Microbiology 142: 1297-1309
19 Zalkin, H. and P. Nyggard. 1999. Biosynthesis of purine nucleotides, pp. 561-579. In F. C. Neidhardt, R. Curtis III, J. L. Ingraham, E. C. C. Lin, K. B.Low, B. Magasanik, W. S. Rennikoff, M. Riley, M. Schaechter, and H. E. Umbarger (eds.), Escherichia coli and Salmonella: Cellular and Molecular Biology, 2nd Ed. American Society for Microbiology, Washington, D.C., U.S.A
20 Blattner, F. R., G. Plunkett III, C. A. Bloch, N. T. Perna, V. Burland, M. Riley, J. Collado-Vides, J. D. Glasner, C. K. Rode, G. F. Mayhew, J. Gregor, N. W. Davis, H. A. Kirkpatrick, M. A. Goeden, D. J. Rose, B. Mau, and Y. Shao. 1997. The complete genome sequence of Escherichia coli K-12. Science 277: 1453-1474
21 Paik, J. E. and B. B. R. Lee. 2003. Isolation of transcription initiation signals from Corynebacterium ammoniagenes and comparison of their gene expression levels in C. ammoniagenes and Escherichia coli. Biotechnol. Lett. 25: 1311-1316
22 Shin, B. S., A. Stein, and H. Zalkin. 1997. Interaction of Bacillus subtilis purine repressor with DNA. J. Bacteriol. 179: 7394-7402
23 Kim, H.-J., J.-S. Park, H.-S. Lee, and Y. Kim. 2004. Utilization of lacZ to isolate regulatory genes from Corynebacterium glutamicum. J. Microbiol. Biotechnol. 12: 336-339
24 Rolfes, R. J. and H. Zalkin. 1990. Autoregulation of Escherichia coli purR requires two control sites downstream of the promoter. J. Bacteriol. 172: 5758-5766
25 Micralv, P., J. E. Bernhard, K. Eva, L. Wolfgang, and S. Hermann. 1996. Promoters from Corynebacterium glutamicum: Cloning, molecular analysis and search for a consensus motif. Microbiology 142: 1297-1309
26 Andersen, L., M. Kilstrup, and J. Neuhard. 1989. Pyrimidine, purine and nitrogen control of cytosine deaminase synthesis in Escherichia coli K12. Involvement of the glnLG and purR genes in the regulation of codA expression. Arch. Microbiol. 152: 115-118
27 Ebbole D. J. and H. Zalkin. 1989. Bacillus subtilis pur operon expression and regulation. J. Bacteriol. 171: 2136-2141
28 Jeon, K. S., G. E. Ji, and I. K. Hwang. 2004. Assay of $\beta$-glucosidase activity of bifidobacteria and the hydrolysis of isoflavone glycosides by Bifidobacterium sp. Int-57 in soymilk fermentation. J. Microbiol. Biotechnol. 12: 8-13
29 Miller, J. H. 1972. Experiments in Molecular Genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., U.S.A
30 He, B., A. Shiau, K. Y. Choi, H. Zalkin, and J. M. Smith. 1990. Genes of the Escherichia coli pur regulon are negatively controlled by a repressor-operator interaction. J. Bacteriol. 172: 4555-4562
31 Han, J. K., S. O. Chung, J. H. Lee, and S. M. Byun. 1997. 6- Mercaptoguanosine resistance is related with purF gene encoding 5-phosphoribosyl-1-pyrophosphate amidotransferase in inosine-5-monophosphate overproducing Brevibacterium ammoniagenes. Biotechnol. Lett. 19: 79-83
32 Saxild, H. H., K. Brunstedt, K. I. Nielsen, H. Jarmer, and P. Nygaard. 2001. Definition of the Bacillus subtilis PurR operator using genetic and bioinformatic tools and expansion of the PurR regulon with glyA, guaC, pbuG, Xpt-pbuX, yqhZ-folD, and pbuO. J. Bacteriol. 183: 6175-6183
33 Zalkin, H. and J. E. Dixon. 1992. De novo purine nucleotide biosynthesis. Prog. Nucl. Acid Res. 42: 259-287
34 Choi, K. Y. and H. Zalkin. 1994. Role of the purine repressor hinge sequences in repressor function. J. Bacteriol. 176: 1767-1772
35 Patek, M., G. Muth, and W. Wohlleben. 2003. Function of Corynebacterium glutamicum promoters in Escherichia coli, Streptomyces lividans, and Bacillus subtilis. J. Biotechnol. 104: 325-334
36 Patek, M., J. Nesvera, A. Guyonvarch, O. Reyes, and G. Leblon. 2003. Promoters of Corynebacterium glutamicum. J. Biotechnol. 104: 311-323
37 Ebbole, D. J. and H. Zalkin. 1987. Cloning and characterization of a 12-gene cluster from Bacillus subtilis encoding nine enzymes for de novo purine nucleotide synthesis. J. Biol. Chem. 262: 8274-8287
38 Rasko, D. A., J. Ravel, O. A. Okstad, E. Helgason, R. Z. Cer, L. Jiang, K. A. Shores, D. E. Fouts, N. J. Tourasse, S. V. Angiuoli, J. Kolonay, W. C. Nelson, A. B. Kolsto, C. M. Fraser, and T. D. Read. 2004. The genome sequence of Bacillus cereus ATCC 10987 reveals metabolic adaptations and a large plasmid related to Bacillus anthracis pXO1. Nucleic Acids Res. 32: 977-988
39 Weng, M., P. Nagy, and H. Zalkin. 1995. Identification of the Bacillus subtilis pur operon repressor. Proc. Natl. Acad. Sci. USA 92: 7455-7459