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. 1984 Sep;159(3):979–985. doi: 10.1128/jb.159.3.979-985.1984

Regulation of the phosphate regulon in Escherichia coli K-12: regulation of the negative regulatory gene phoU and identification of the gene product.

A Nakata, M Amemura, H Shinagawa
PMCID: PMC215756  PMID: 6090402

Abstract

The phoU gene is one of the negative regulatory genes of the pho regulon of Escherichia coli. The DNA fragment carrying phoU has been cloned on pBR322 (Amemura et al., J. Bacteriol. 152:692-701, 1982). Further subcloning, Tn1000 insertion inactivation, and complementation tests localized the phoU gene within a 1.1-kilobase region on the cloned DNA fragment. The gene product of phoU was identified by the maxicell method as a protein with an approximate molecular weight of 27,000. A hybrid plasmid that contains a phoU'-lac'Z fused gene was constructed in vitro. This plasmid enabled us to study phoU gene expression by measuring the beta-galactosidase level in the cells. The plasmid was introduced into various regulatory mutants related to the pho regulon, and phoU gene expression in these strains was studied under limited and excess phosphate conditions. It was found that phoU is expressed at a higher level when the cells are cultured under the excess phosphate condition. The higher phoU expression was observed in a phoB mutant and a phoR-phoM double mutant. The implications of these findings for the regulation of pho genes are discussed.

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Selected References

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  1. Amemura M., Shinagawa H., Makino K., Otsuji N., Nakata A. Cloning of and complementation tests with alkaline phosphatase regulatory genes (phoS and phoT) of Escherichia coli. J Bacteriol. 1982 Nov;152(2):692–701. doi: 10.1128/jb.152.2.692-701.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bachmann B. J. Linkage map of Escherichia coli K-12, edition 7. Microbiol Rev. 1983 Jun;47(2):180–230. doi: 10.1128/mr.47.2.180-230.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bassford P. J., Jr, Silhavy T. J., Beckwith J. R. Use of gene fusion to study secretion of maltose-binding protein into Escherichia coli periplasm. J Bacteriol. 1979 Jul;139(1):19–31. doi: 10.1128/jb.139.1.19-31.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Casadaban M. J., Chou J., Cohen S. N. In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals. J Bacteriol. 1980 Aug;143(2):971–980. doi: 10.1128/jb.143.2.971-980.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cox G. B., Rosenberg H., Downie J. A., Silver S. Genetic analysis of mutants affected in the Pst inorganic phosphate transport system. J Bacteriol. 1981 Oct;148(1):1–9. doi: 10.1128/jb.148.1.1-9.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Magota K., Otsuji N., Miki T., Horiuchi T., Tsunasawa S., Kondo J., Sakiyama F., Amemura M., Morita T., Shinagawa H. Nucleotide sequence of the phoS gene, the structural gene for the phosphate-binding protein of Escherichia coli. J Bacteriol. 1984 Mar;157(3):909–917. doi: 10.1128/jb.157.3.909-917.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Manis J. J., Kline B. C. Restriction endonuclease mapping and mutagenesis of the F sex factor replication region. Mol Gen Genet. 1977 Apr 29;152(3):175–182. doi: 10.1007/BF00268815. [DOI] [PubMed] [Google Scholar]
  9. Morita T., Amemura M., Makino K., Shinagawa H., Magota K., Otsuji N., Nakata A. Hyperproduction of phosphate-binding protein, phoS, and pre-phoS proteins in Escherichia coli carrying a cloned phoS gene. Eur J Biochem. 1983 Feb 15;130(3):427–435. doi: 10.1111/j.1432-1033.1983.tb07169.x. [DOI] [PubMed] [Google Scholar]
  10. Nakata A., Peterson G. R., Brooks E. L., Rothman F. G. Location and orientation of the phoA locus on the Escherichia coli K-12 linkage map. J Bacteriol. 1971 Sep;107(3):683–689. doi: 10.1128/jb.107.3.683-689.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Rosenberg H., Gerdes R. G., Chegwidden K. Two systems for the uptake of phosphate in Escherichia coli. J Bacteriol. 1977 Aug;131(2):505–511. doi: 10.1128/jb.131.2.505-511.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sancar A., Wharton R. P., Seltzer S., Kacinski B. M., Clarke N. D., Rupp W. D. Identification of the uvrA gene product. J Mol Biol. 1981 May 5;148(1):45–62. doi: 10.1016/0022-2836(81)90234-5. [DOI] [PubMed] [Google Scholar]
  13. Shinagawa H., Makino K., Nakata A. Regulation of the pho regulon in Escherichia coli K-12. Genetic and physiological regulation of the positive regulatory gene phoB. J Mol Biol. 1983 Aug 15;168(3):477–488. doi: 10.1016/s0022-2836(83)80297-6. [DOI] [PubMed] [Google Scholar]
  14. Silhavy T. J., Shuman H. A., Beckwith J., Schwartz M. Use of gene fusions to study outer membrane protein localization in Escherichia coli. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5411–5415. doi: 10.1073/pnas.74.12.5411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sprague G. F., Jr, Bell R. M., Cronan J. E., Jr A mutant of Escherichia coli auxotrophic for organic phosphates: evidence for two defects in inorganic phosphate transport. Mol Gen Genet. 1975 Dec 30;143(1):71–77. doi: 10.1007/BF00269422. [DOI] [PubMed] [Google Scholar]
  16. Surin B. P., Jans D. A., Fimmel A. L., Shaw D. C., Cox G. B., Rosenberg H. Structural gene for the phosphate-repressible phosphate-binding protein of Escherichia coli has its own promoter: complete nucleotide sequence of the phoS gene. J Bacteriol. 1984 Mar;157(3):772–778. doi: 10.1128/jb.157.3.772-778.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tommassen J., Lugtenberg B. PHO-regulon of Escherichia coli K12: a minireview. Ann Microbiol (Paris) 1982 Mar-Apr;133(2):243–249. [PubMed] [Google Scholar]
  18. Wanner B. L., Latterell P. Mutants affected in alkaline phosphatase, expression: evidence for multiple positive regulators of the phosphate regulon in Escherichia coli. Genetics. 1980 Oct;96(2):353–366. doi: 10.1093/genetics/96.2.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Willsky G. R., Bennett R. L., Malamy M. H. Inorganic phosphate transport in Escherichia coli: involvement of two genes which play a role in alkaline phosphatase regulation. J Bacteriol. 1973 Feb;113(2):529–539. doi: 10.1128/jb.113.2.529-539.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Zuckier G., Ingenito E., Torriani A. Pleiotropic effects of alkaline phosphatase regulatory mutations phoB and phoT on anaerobic growth of and polyphosphate synthesis in Escherichia coli. J Bacteriol. 1980 Aug;143(2):934–941. doi: 10.1128/jb.143.2.934-941.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Zuckier G., Torriani A. Genetic and physiological tests of three phosphate-specific transport mutants of Escherichia coli. J Bacteriol. 1981 Mar;145(3):1249–1256. doi: 10.1128/jb.145.3.1249-1256.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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