Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Feb;149(2):434–439. doi: 10.1128/jb.149.2.434-439.1982

Signal sequence of alkaline phosphatase of Escherichia coli.

H Inouye, W Barnes, J Beckwith
PMCID: PMC216526  PMID: 7035431

Abstract

The amino acid sequence of the signal sequence of phoA was determined by DNA sequencing by using the dideoxy chain termination technique (Sanger et al., Proc. Natl. Acad. Sci. U.S.A. 74:5463-5467, 1977). The template used was single-stranded DNA obtained from M13 on f1 phage derivatives carrying phoA, constructed by in vitro recombination. The results confirm the sequence of the first five amino acids determined by Sarthy et al. (J. Bacteriol. 139:932-939, 1979) and extend the sequence in the same reading frame into the amino terminal region of the mature alkaline phosphatase (Bradshaw et al., Proc. Natl. Acad. Sci. U.S.A., 78:3473-3477, 1981). As was predicted (Inouye and Beckwith, Proc. Natl. Acad. Sci. U.S.A. 74:1440-1444, 1977), the signal sequence was highly hydrophobic. The alteration of DNA sequence was identified for a promoter mutation that results in the expression of phoA independent of the positive control gene phoB and in insensitivity to high phosphate.

Full text

PDF
438

Images in this article

437Image
on p.437

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barnes W. M. Construction of an M13 histidine-transducing phage: a single-stranded cloning vehicle with one EcoRI site. Gene. 1979 Feb;5(2):127–139. doi: 10.1016/0378-1119(79)90098-2. [DOI] [PubMed] [Google Scholar]
  3. Barnes W. M. DNA sequence from the histidine operon control region: seven histidine codons in a row. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4281–4285. doi: 10.1073/pnas.75.9.4281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Barnes W. M. Plasmid detection and sizing in single colony lysates. Science. 1977 Jan 28;195(4276):393–394. doi: 10.1126/science.318764. [DOI] [PubMed] [Google Scholar]
  5. Bradshaw R. A., Cancedda F., Ericsson L. H., Neumann P. A., Piccoli S. P., Schlesinger M. J., Shriefer K., Walsh K. A. Amino acid sequence of Escherichia coli alkaline phosphatase. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3473–3477. doi: 10.1073/pnas.78.6.3473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brickman E., Beckwith J. Analysis of the regulation of Escherichia coli alkaline phosphatase synthesis using deletions and phi80 transducing phages. J Mol Biol. 1975 Aug 5;96(2):307–316. doi: 10.1016/0022-2836(75)90350-2. [DOI] [PubMed] [Google Scholar]
  7. Chang C. N., Inouye H., Model P., Beckwith J. Processing of alkaline phosphatase precursor to the mature enzyme by an Escherichia coli inner membrane preparation. J Bacteriol. 1980 May;142(2):726–728. doi: 10.1128/jb.142.2.726-728.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen S. N., Chang A. C., Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci U S A. 1972 Aug;69(8):2110–2114. doi: 10.1073/pnas.69.8.2110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Farabaugh P. J. Sequence of the lacI gene. Nature. 1978 Aug 24;274(5673):765–769. doi: 10.1038/274765a0. [DOI] [PubMed] [Google Scholar]
  10. Inouye H., Beckwith J. Synthesis and processing of an Escherichia coli alkaline phosphatase precursor in vitro. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1440–1444. doi: 10.1073/pnas.74.4.1440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Inouye H., Michaelis S., Wright A., Beckwith J. Cloning and restriction mapping of the alkaline phosphatase structural gene (phoA) of Escherichia coli and generation of deletion mutants in vitro. J Bacteriol. 1981 May;146(2):668–675. doi: 10.1128/jb.146.2.668-675.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ito K., Beckwith J. R. Role of the mature protein sequence of maltose-binding protein in its secretion across the E. coli cytoplasmic membrane. Cell. 1981 Jul;25(1):143–150. doi: 10.1016/0092-8674(81)90238-5. [DOI] [PubMed] [Google Scholar]
  13. Jeppesen P. G. A method for separating DNA fragments by electrophoresis in polyacrylamide concentration gradient slab gels. Anal Biochem. 1974 Mar;58(1):195–207. doi: 10.1016/0003-2697(74)90458-8. [DOI] [PubMed] [Google Scholar]
  14. MALAMY M., HORECKER B. L. The localization of alkaline phosphatase in E. coli K12. Biochem Biophys Res Commun. 1961 Jun 2;5:104–108. doi: 10.1016/0006-291x(61)90020-1. [DOI] [PubMed] [Google Scholar]
  15. Post L. E., Arfsten A. E., Davis G. R., Nomura M. DNA sequence of the promoter region for the alpha ribosomal protein operon in Escherichia coli. J Biol Chem. 1980 May 25;255(10):4653–4659. [PubMed] [Google Scholar]
  16. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sarthy A., Fowler A., Zabin I., Beckwith J. Use of gene fusions to determine a partial signal sequence of alkaline phosphatase. J Bacteriol. 1979 Sep;139(3):932–939. doi: 10.1128/jb.139.3.932-939.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sarthy A., Michaelis S., Beckwith J. Use of gene fusions to determine the orientation of gene phoA on the Escherichia coli chromosome. J Bacteriol. 1981 Jan;145(1):293–298. doi: 10.1128/jb.145.1.293-298.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Shine J., Dalgarno L. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342–1346. doi: 10.1073/pnas.71.4.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sutcliffe J. G. Nucleotide sequence of the ampicillin resistance gene of Escherichia coli plasmid pBR322. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3737–3741. doi: 10.1073/pnas.75.8.3737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Wanner B. L., Sarthy A., Beckwith J. Escherichia coli pleiotropic mutant that reduces amounts of several periplasmic and outer membrane proteins. J Bacteriol. 1979 Oct;140(1):229–239. doi: 10.1128/jb.140.1.229-239.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Yokota T., Sugisaki H., Takanami M., Kaziro Y. The nucleotide sequence of the cloned tufA gene of Escherichia coli. Gene. 1980 Dec;12(1-2):25–31. doi: 10.1016/0378-1119(80)90012-8. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES