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. 1984 Mar;157(3):909–917. doi: 10.1128/jb.157.3.909-917.1984

Nucleotide sequence of the phoS gene, the structural gene for the phosphate-binding protein of Escherichia coli.

K Magota, N Otsuji, T Miki, T Horiuchi, S Tsunasawa, J Kondo, F Sakiyama, M Amemura, T Morita, H Shinagawa, et al.
PMCID: PMC215345  PMID: 6365894

Abstract

phoS is the structural gene for the phosphate-binding protein, which is localized in periplasm and involved in active transport of phosphate in Escherichia coli. It is also a negative regulatory gene for the pho regulon, and the gene expression is inducible by phosphate starvation. The complete nucleotide sequence of the phoS gene was determined by the method of Maxam and Gilbert (A. M. Maxam and W. Gilbert, Methods Enzymol. 65:499-560, 1980). The amino acid sequences at the amino termini of the pre-PhoS and PhoS proteins and at the carboxy terminus of the PhoS protein were determined by using the purified proteins. Furthermore, the amino acid sequence of enzymatically digested peptide fragments of the PhoS protein was determined. The combined data established the nucleotide sequence of the coding region and the amino acid sequence of the pre-PhoS and the PhoS proteins. The pre-PhoS protein contains an extension of peptide composed of 25 amino acid residues at the amino terminus of the PhoS protein, which has the general characteristics of a signal peptide. The mature PhoS protein is composed of 321 amino acid residues, with a calculated molecular weight of 34,422, and lacks the disulfide bond and methionine. The regulatory region of phoS contains a characteristic Shine-Dalgarno sequence at an appropriate position preceding the translational initiation site, as well as three possible Pribnow boxes and one -35 sequence. the nucleotide sequence of the regulatory region of phoS was compared with those of phoA and phoE, the genes constituting the pho regulon.

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Selected References

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  1. Akeroyd R., Lange L. G., Westerman J., Wirtz K. W. Modification of the phosphatidylcholine-transfer protein from bovine liver with butanedione and phenylglyoxal. Evidence for one essential arginine residue. Eur J Biochem. 1981 Dec;121(1):77–81. doi: 10.1111/j.1432-1033.1981.tb06432.x. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Aono H., Otsuji N. Genetic mapping of regulator gene phoS for alkaline phosphatase in Escherichia coli. J Bacteriol. 1968 Mar;95(3):1182–1183. doi: 10.1128/jb.95.3.1182-1183.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Argast M., Boos W. Co-regulation in Escherichia coli of a novel transport system for sn-glycerol-3-phosphate and outer membrane protein Ic (e, E) with alkaline phosphatase and phosphate-binding protein. J Bacteriol. 1980 Jul;143(1):142–150. doi: 10.1128/jb.143.1.142-150.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Bennett R. L., Malamy M. H. Arsenate resistant mutants of Escherichia coli and phosphate transport. Biochem Biophys Res Commun. 1970 Jul 27;40(2):496–503. doi: 10.1016/0006-291x(70)91036-3. [DOI] [PubMed] [Google Scholar]
  7. Borer P. N., Dengler B., Tinoco I., Jr, Uhlenbeck O. C. Stability of ribonucleic acid double-stranded helices. J Mol Biol. 1974 Jul 15;86(4):843–853. doi: 10.1016/0022-2836(74)90357-x. [DOI] [PubMed] [Google Scholar]
  8. Bracha M., Yagil E. A ne type of alkaline phosphatase-negative mutants in Escherichia coli K12. Mol Gen Genet. 1973 Mar 27;122(1):53–60. doi: 10.1007/BF00337973. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Dianoux A. C., Vignais P. V., Tsugita A. Partial proteolysis of the natural ATPase inhibitor from beef heart mitochondria. Isolation and characterization of an active cleavage product. FEBS Lett. 1981 Jul 20;130(1):119–123. doi: 10.1016/0014-5793(81)80678-3. [DOI] [PubMed] [Google Scholar]
  11. ECHOLS H., GAREN A., GAREN S., TORRIANI A. Genetic control of repression of alkaline phosphatase in E. coli. J Mol Biol. 1961 Aug;3:425–438. doi: 10.1016/s0022-2836(61)80055-7. [DOI] [PubMed] [Google Scholar]
  12. Emr S. D., Hall M. N., Silhavy T. J. A mechanism of protein localization: the signal hypothesis and bacteria. J Cell Biol. 1980 Sep;86(3):701–711. doi: 10.1083/jcb.86.3.701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. GAREN A., ECHOLS H. Genetic control of induction of alkaline phosphatase synthesis in E. coli. Proc Natl Acad Sci U S A. 1962 Aug;48:1398–1402. doi: 10.1073/pnas.48.8.1398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. GAREN A., ECHOLS H. Properties of two regulating genes for alkaline phosphatase. J Bacteriol. 1962 Feb;83:297–300. doi: 10.1128/jb.83.2.297-300.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. GAREN A., OTSUJI N. ISOLATION OF A PROTEIN SPECIFIED BY A REGULATOR GENE. J Mol Biol. 1964 Jun;8:841–852. doi: 10.1016/s0022-2836(64)80165-0. [DOI] [PubMed] [Google Scholar]
  16. Gerdes R. G., Rosenberg H. The relationship between the phosphate-binding protein and a regulator gene product from Escherichia coli. Biochim Biophys Acta. 1974 May 10;351(1):77–86. doi: 10.1016/0005-2795(74)90066-x. [DOI] [PubMed] [Google Scholar]
  17. Hawley D. K., McClure W. R. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res. 1983 Apr 25;11(8):2237–2255. doi: 10.1093/nar/11.8.2237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Inouye H., Barnes W., Beckwith J. Signal sequence of alkaline phosphatase of Escherichia coli. J Bacteriol. 1982 Feb;149(2):434–439. doi: 10.1128/jb.149.2.434-439.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kikuchi Y., Yoda K., Yamasaki M., Tamura G. The nucleotide sequence of the promoter and the amino-terminal region of alkaline phosphatase structural gene (phoA) of Escherichia coli. Nucleic Acids Res. 1981 Nov 11;9(21):5671–5678. doi: 10.1093/nar/9.21.5671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kreuzer K., Pratt C., Torriani A. Genetic analysis of regulatory mutants of alkaline phosphatase of E. coli. Genetics. 1975 Nov;81(3):459–468. doi: 10.1093/genetics/81.3.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kupersztoch-Portnoy Y. M., Lovett M. A., Helinski D. R. Strand and site specificity of the relaxation event for the relaxation complex of the antibiotic resistance plasmid R6K. Biochemistry. 1974 Dec 31;13(27):5484–5490. doi: 10.1021/bi00724a005. [DOI] [PubMed] [Google Scholar]
  22. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  23. Levitz R., Bittan R., Yagil E. Complementation tests between alkaline phosphatase-constitutive mutants (phoS and phoT) of Escherichia coli. J Bacteriol. 1981 Mar;145(3):1432–1435. doi: 10.1128/jb.145.3.1432-1435.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  25. Medveczky N., Rosenberg H. The binding and release of phosphate by a protein isolated from Escherichia coli. Biochim Biophys Acta. 1969 Nov 18;192(2):369–371. doi: 10.1016/0304-4165(69)90382-1. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Morkin E., Flink I. L., Banerjee S. K. Phenylglyoxal modification of cardiac myosin S-1. Evidence for essential arginine residues at the active site. J Biol Chem. 1979 Dec 25;254(24):12647–12652. [PubMed] [Google Scholar]
  28. Overbeeke N., Bergmans H., van Mansfeld F., Lugtenberg B. Complete nucleotide sequence of phoE, the structural gene for the phosphate limitation inducible outer membrane pore protein of Escherichia coli K12. J Mol Biol. 1983 Feb 5;163(4):513–532. doi: 10.1016/0022-2836(83)90110-9. [DOI] [PubMed] [Google Scholar]
  29. Overbeeke N., Lugtenberg B. Expression of outer membrane protein e of Escherichia coli K12 by phosphate limitation. FEBS Lett. 1980 Apr 7;112(2):229–232. doi: 10.1016/0014-5793(80)80186-4. [DOI] [PubMed] [Google Scholar]
  30. Pratt C., Torriani A. Complementation test between alkaline phosphatase regulatory mutations phoB and phoRc in Escherichia coli. Genetics. 1977 Feb;85(2):203–208. doi: 10.1093/genetics/85.2.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  33. Schrijen J. J., Luyben W. A., De Pont J. J., Bonting S. L. Studies on (K+ + H+)-ATPase. I. Essential arginine residue in its substrate binding center. Biochim Biophys Acta. 1980 Apr 10;597(2):331–344. doi: 10.1016/0005-2736(80)90110-8. [DOI] [PubMed] [Google Scholar]
  34. 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]
  35. Silhavy T. J., Casadaban M. J., Shuman H. A., Beckwith J. R. Conversion of beta-galactosidase to a membrane-bound state by gene fusion. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3423–3427. doi: 10.1073/pnas.73.10.3423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Simpson R. J., Neuberger M. R., Liu T. Y. Complete amino acid analysis of proteins from a single hydrolysate. J Biol Chem. 1976 Apr 10;251(7):1936–1940. [PubMed] [Google Scholar]
  37. 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]
  38. Staden R. Further procedures for sequence analysis by computer. Nucleic Acids Res. 1978 Mar;5(3):1013–1016. doi: 10.1093/nar/5.3.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tinoco I., Jr, Borer P. N., Dengler B., Levin M. D., Uhlenbeck O. C., Crothers D. M., Bralla J. Improved estimation of secondary structure in ribonucleic acids. Nat New Biol. 1973 Nov 14;246(150):40–41. doi: 10.1038/newbio246040a0. [DOI] [PubMed] [Google Scholar]
  40. Tommassen J., Lugtenberg B. Outer membrane protein e of Escherichia coli K-12 is co-regulated with alkaline phosphatase. J Bacteriol. 1980 Jul;143(1):151–157. doi: 10.1128/jb.143.1.151-157.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. 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]
  42. 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]
  43. Willsky G. R., Malamy M. H. Control of the synthesis of alkaline phosphatase and the phosphate-binding protein in Escherichia coli. J Bacteriol. 1976 Jul;127(1):595–609. doi: 10.1128/jb.127.1.595-609.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Zimmerman C. L., Appella E., Pisano J. J. Rapid analysis of amino acid phenylthiohydantoins by high-performance liquid chromatography. Anal Biochem. 1977 Feb;77(2):569–573. doi: 10.1016/0003-2697(77)90276-7. [DOI] [PubMed] [Google Scholar]
  45. 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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