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. 1984 Aug 10;12(15):6337–6355. doi: 10.1093/nar/12.15.6337

Analysis of the Escherichia coli proBA locus by DNA and protein sequencing.

A H Deutch, K E Rushlow, C J Smith
PMCID: PMC320076  PMID: 6089111

Abstract

A 2.9 kb DNA fragment carrying the Escherichia coli proBA region, which encodes the first two enzymes of the proline biosynthetic pathway, was subcloned onto an expression plasmid carrying both the bacteriophage lambda PL promoter (lambda PL) and the lambda gene encoding a thermolabile cI repressor protein (cI857). Derepression of the lambda PL promoter by thermal inactivation of the cI857 repressor protein resulted in the simultaneous overproduction of the proB (gamma-glutamyl kinase) and proA (gamma-glutamyl phosphate reductase) gene products. Nucleotide sequence analysis of the proBA locus allowed gene assignments consistent with the NH2 and COOH-terminal analyses and amino acid compositions of homogeneous preparations of the proB and proA proteins. The contiguous nature of the proB and proA genes suggests that the two genes constitute an operon in which proB precedes proA.

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Selected References

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

  1. Alff-Steinberger C. Evidence for a coding pattern on the non-coding strand of the E. coli genome. Nucleic Acids Res. 1984 Mar 12;12(5):2235–2241. doi: 10.1093/nar/12.5.2235. [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. Baich A., Pierson D. J. Control of proline synthesis in Escherichia coli. Biochim Biophys Acta. 1965 Jul 8;104(2):397–404. doi: 10.1016/0304-4165(65)90345-4. [DOI] [PubMed] [Google Scholar]
  4. Baich A. Proline synthesis in Escherichia coli. A proline-inhibitable glutamic acid kinase. Biochim Biophys Acta. 1969 Dec 30;192(3):462–467. doi: 10.1016/0304-4165(69)90395-x. [DOI] [PubMed] [Google Scholar]
  5. Baich A. The biosynthesis of proline in Escherichia coli: phosphate-dependent glutamate -semialdehyde dehydrogenase (NADP), the second enzyme in the pathway. Biochim Biophys Acta. 1971 Jul 20;244(1):129–134. doi: 10.1016/0304-4165(71)90129-2. [DOI] [PubMed] [Google Scholar]
  6. Broda P. Modified map positions for lac and the pro markers in Escherichia coli K-12. J Bacteriol. 1974 Feb;117(2):741–746. doi: 10.1128/jb.117.2.741-746.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Clarke L., Carbon J. A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell. 1976 Sep;9(1):91–99. doi: 10.1016/0092-8674(76)90055-6. [DOI] [PubMed] [Google Scholar]
  8. Condamine H. Mutants des voies de biosynthèse et de dégradation de la proline chez E. coli K 12. Ann Inst Pasteur (Paris) 1971 Jan;120(1):9–22. [PubMed] [Google Scholar]
  9. Deutch A. H., Smith C. J., Rushlow K. E., Kretschmer P. J. Escherichia coli delta 1-pyrroline-5-carboxylate reductase: gene sequence, protein overproduction and purification. Nucleic Acids Res. 1982 Dec 11;10(23):7701–7714. doi: 10.1093/nar/10.23.7701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Havekes L. M., Lugtenberg B. J., Hoekstra W. P. Conjugation deficient E. coli K12 F- mutants with heptose-less lipopolysaccharide. Mol Gen Genet. 1976 Jul 5;146(1):43–50. doi: 10.1007/BF00267981. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Hayzer D. J., Leisinger T. Proline biosynthesis in Escherichia coli. Purification and characterisation of glutamate-semialdehyde dehydrogenase. Eur J Biochem. 1982 Jan;121(3):561–565. doi: 10.1111/j.1432-1033.1982.tb05823.x. [DOI] [PubMed] [Google Scholar]
  13. Hayzer D. J., Leisinger T. The gene-enzyme relationships of proline biosynthesis in Escherichia coli. J Gen Microbiol. 1980 Jun;118(2):287–293. doi: 10.1099/00221287-118-2-287. [DOI] [PubMed] [Google Scholar]
  14. Hayzer D. J. Sub-cloning of the wild-type proAB region of the Escherichia coli genome. J Gen Microbiol. 1983 Oct;129(10):3215–3225. doi: 10.1099/00221287-129-10-3215. [DOI] [PubMed] [Google Scholar]
  15. Hoopes B. C., McClure W. R. Studies on the selectivity of DNA precipitation by spermine. Nucleic Acids Res. 1981 Oct 24;9(20):5493–5504. doi: 10.1093/nar/9.20.5493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Karp D. R., Parker K. L., Shreffler D. C., Capra J. D. Characterization of the murine C4 precursor (pro-C4): evidence that the carboxy-terminal subunit is the C4 gamma-chain. J Immunol. 1981 May;126(5):2060–2061. [PubMed] [Google Scholar]
  17. Klapper D. G., Wilde C. E., 3rd, Capra J. D. Automated amino acid sequence of small peptides utilizing Polybrene. Anal Biochem. 1978 Mar;85(1):126–131. doi: 10.1016/0003-2697(78)90282-8. [DOI] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Le Rudulier D., Strom A. R., Dandekar A. M., Smith L. T., Valentine R. C. Molecular biology of osmoregulation. Science. 1984 Jun 8;224(4653):1064–1068. doi: 10.1126/science.224.4653.1064. [DOI] [PubMed] [Google Scholar]
  20. Mahan M. J., Csonka L. N. Genetic analysis of the proBA genes of Salmonella typhimurium: physical and genetic analyses of the cloned proB+ A+ genes of Escherichia coli and of a mutant allele that confers proline overproduction and enhanced osmotolerance. J Bacteriol. 1983 Dec;156(3):1249–1262. doi: 10.1128/jb.156.3.1249-1262.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  23. Oppenheim D. S., Yanofsky C. Translational coupling during expression of the tryptophan operon of Escherichia coli. Genetics. 1980 Aug;95(4):785–795. doi: 10.1093/genetics/95.4.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Pinkham J. L., Platt T., Enquist L. W., Weisberg R. A. The secondary attachment site for bacteriophage lambda in the proA/B gene of Escherichia coli. J Mol Biol. 1980 Dec 25;144(4):587–592. doi: 10.1016/0022-2836(80)90339-3. [DOI] [PubMed] [Google Scholar]
  26. Roberts L. M., Reeve E. C. Two mutations giving low-level streptomycin resistance in Escherichia coli K 12. Genet Res. 1970 Dec;16(3):359–365. doi: 10.1017/s0016672300002640. [DOI] [PubMed] [Google Scholar]
  27. Rossi J. J., Vender J., Berg C. M., Coleman W. H. Partial purification and some properties of delta1-pyrroline-5-carboxylate reductase from Escherichia coli. J Bacteriol. 1977 Jan;129(1):108–114. doi: 10.1128/jb.129.1.108-114.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sanger F., Coulson A. R., Barrell B. G., Smith A. J., Roe B. A. Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol. 1980 Oct 25;143(2):161–178. doi: 10.1016/0022-2836(80)90196-5. [DOI] [PubMed] [Google Scholar]
  29. Shimada K., Weisberg R. A., Gottesman M. E. Prophage lambda at unusual chromosomal locations. I. Location of the secondary attachment sites and the properties of the lysogens. J Mol Biol. 1972 Feb 14;63(3):483–503. doi: 10.1016/0022-2836(72)90443-3. [DOI] [PubMed] [Google Scholar]
  30. Smith C. J., Deutch A. H., Rushlow K. E. Purification and characteristics of a gamma-glutamyl kinase involved in Escherichia coli proline biosynthesis. J Bacteriol. 1984 Feb;157(2):545–551. doi: 10.1128/jb.157.2.545-551.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Smith G. R., Kunes S. M., Schultz D. W., Taylor A., Triman K. L. Structure of chi hotspots of generalized recombination. Cell. 1981 May;24(2):429–436. doi: 10.1016/0092-8674(81)90333-0. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Tristram H., Thurston C. F. Control of proline biosynthesis by proline and proline analogues. Nature. 1966 Oct 1;212(5057):74–75. doi: 10.1038/212074a0. [DOI] [PubMed] [Google Scholar]
  34. Williams I., Frank L. Improved chemical synthesis and enzymatic assay of delta-1-pyrroline-5-carboxylic acid. Anal Biochem. 1975 Mar;64(1):85–97. doi: 10.1016/0003-2697(75)90408-x. [DOI] [PubMed] [Google Scholar]

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