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. 1987 Oct;169(10):4716–4721. doi: 10.1128/jb.169.10.4716-4721.1987

Structural and functional analysis of a cloned segment of Escherichia coli DNA that specifies proteins of a C4 pathway of serine biosynthesis.

P D Ravnikar 1, R L Somerville 1
PMCID: PMC213845  PMID: 2820941

Abstract

The plasmid pDR121 is a pBR322 derivative that contains a 3.7-kilobase-pair EcoRI fragment of DNA from the 81.2-min region of the Escherichia coli chromosome. The genomic insert encodes threonine dehydrogenase and at least one other protein. Several physical and kinetic properties of threonine dehydrogenase, overproduced in cells harboring pDR121, are identical to those of pure threonine dehydrogenase from a haploid mutant of E. coli K-12 that produces this enzyme constitutively. Tester strains with serB or glyA mutations harboring pDR121 are prototrophs. The ability to confer prototrophy on such tester strains is associated with elevated levels of threonine dehydrogenase. The functional roles of various segments of the 3.7-kilobase-pair insert of pDR121 were analyzed by constructing specific deletions and insertions. Certain subclones retained the ability to specify threonine dehydrogenase without conferring prototrophy on tester strains. This suggests that at least one other protein encoded within pDR121 plays an essential role in the conversion of threonine to serine.

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Selected References

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

  1. 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]
  2. Bird M. I., Nunn P. B., Lord L. A. Formation of glycine and aminoacetone from L-threonine by rat liver mitochondria. Biochim Biophys Acta. 1984 Nov 28;802(2):229–236. doi: 10.1016/0304-4165(84)90166-1. [DOI] [PubMed] [Google Scholar]
  3. Bogosian G., Bertrand K., Somerville R. Trp repressor protein controls its own structural gene. J Mol Biol. 1981 Jul 15;149(4):821–825. doi: 10.1016/0022-2836(81)90361-2. [DOI] [PubMed] [Google Scholar]
  4. Boylan S. A., Dekker E. E. L-threonine dehydrogenase. Purification and properties of the homogeneous enzyme from Escherichia coli K-12. J Biol Chem. 1981 Feb 25;256(4):1809–1815. [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Brown T. D., Jones-Mortimer M. C., Kornberg H. L. The enzymic interconversion of acetate and acetyl-coenzyme A in Escherichia coli. J Gen Microbiol. 1977 Oct;102(2):327–336. doi: 10.1099/00221287-102-2-327. [DOI] [PubMed] [Google Scholar]
  7. Chan T. T., Newman E. B. Threonine as a carbon source for Escherichia coli. J Bacteriol. 1981 Mar;145(3):1150–1153. doi: 10.1128/jb.145.3.1150-1153.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Clarke S. J., Low B., Konigsberg W. H. Close linkage of the genes serC (for phosphohydroxy pyruvate transaminase) and serS (for seryl-transfer ribonucleic acid synthetase) in Escherichia coli K-12. J Bacteriol. 1973 Mar;113(3):1091–1095. doi: 10.1128/jb.113.3.1091-1095.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Close T. J., Rodriguez R. L. Construction and characterization of the chloramphenicol-resistance gene cartridge: a new approach to the transcriptional mapping of extrachromosomal elements. Gene. 1982 Dec;20(2):305–316. doi: 10.1016/0378-1119(82)90048-8. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Duncan K., Coggins J. R. The serC-aro A operon of Escherichia coli. A mixed function operon encoding enzymes from two different amino acid biosynthetic pathways. Biochem J. 1986 Feb 15;234(1):49–57. doi: 10.1042/bj2340049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Green M. L., Elliott W. H. The enzymic formation of aminoacetone from threonine and its further metabolism. Biochem J. 1964 Sep;92(3):537–549. doi: 10.1042/bj0920537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. HARTSHORNE D., GREENBERG D. M. STUDIES ON LIVER THREONINE DEHYDROGENASE. Arch Biochem Biophys. 1964 Apr;105:173–178. doi: 10.1016/0003-9861(64)90250-4. [DOI] [PubMed] [Google Scholar]
  14. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  16. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  17. Lathe R., Kieny M. P., Skory S., Lecocq J. P. Linker tailing: unphosphorylated linker oligonucleotides for joining DNA termini. DNA. 1984;3(2):173–182. doi: 10.1089/dna.1984.3.173. [DOI] [PubMed] [Google Scholar]
  18. Lessie T. G., Whiteley H. R. Properties of threonine deaminase from a bacterium able to use threonine as sole source of carbon. J Bacteriol. 1969 Nov;100(2):878–889. doi: 10.1128/jb.100.2.878-889.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McGilvray D., Morris J. G. Utilization of L-threonine by a species of Arthrobacter. A novel catabolic role for "aminoacetone synthase". Biochem J. 1969 May;112(5):657–671. doi: 10.1042/bj1120657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. NEUBERGER A., TAIT G. H. Production of aminoacetone by Rhodopseudomonas spheroides. Biochem J. 1962 Aug;84:317–328. doi: 10.1042/bj0840317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Neuwald A. F., Stauffer G. V. DNA sequence and characterization of the Escherichia coli serB gene. Nucleic Acids Res. 1985 Oct 11;13(19):7025–7039. doi: 10.1093/nar/13.19.7025. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Newman E. B., Kapoor V., Potter R. Role of L-threonine dehydrogenase in the catabolism of threonine and synthesis of glycine by Escherichia coli. J Bacteriol. 1976 Jun;126(3):1245–1249. doi: 10.1128/jb.126.3.1245-1249.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. PIZER L. I. GLYCINE SYNTHESIS AND METABOLISM IN ESCHERICHIA COLI. J Bacteriol. 1965 Apr;89:1145–1150. doi: 10.1128/jb.89.4.1145-1150.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. PIZER L. I. THE PATHWAY AND CONTROL OF SERINE BIOSYNTHESIS IN ESCHERICHIA COLI. J Biol Chem. 1963 Dec;238:3934–3944. [PubMed] [Google Scholar]
  25. Plamann M. D., Stauffer L. T., Urbanowski M. L., Stauffer G. V. Complete nucleotide sequence of the E. coli glyA gene. Nucleic Acids Res. 1983 Apr 11;11(7):2065–2075. doi: 10.1093/nar/11.7.2065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Prentki P., Karch F., Iida S., Meyer J. The plasmid cloning vector pBR325 contains a 482 base-pair-long inverted duplication. Gene. 1981 Sep;14(4):289–299. doi: 10.1016/0378-1119(81)90161-x. [DOI] [PubMed] [Google Scholar]
  27. Ravnikar P. D., Somerville R. L. Genetic characterization of a highly efficient alternate pathway of serine biosynthesis in Escherichia coli. J Bacteriol. 1987 Jun;169(6):2611–2617. doi: 10.1128/jb.169.6.2611-2617.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ravnikar P. D., Somerville R. L. Localization of the structural gene for threonine dehydrogenase in Escherichia coli. J Bacteriol. 1986 Oct;168(1):434–436. doi: 10.1128/jb.168.1.434-436.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ray M., Ray S. L-Threonine dehydrogenase from goat liver. Feedback inhibition by methylglyoxal. J Biol Chem. 1985 May 25;260(10):5913–5918. [PubMed] [Google Scholar]
  30. Soberon X., Covarrubias L., Bolivar F. Construction and characterization of new cloning vehicles. IV. Deletion derivatives of pBR322 and pBR325. Gene. 1980 May;9(3-4):287–305. doi: 10.1016/0378-1119(90)90328-o. [DOI] [PubMed] [Google Scholar]
  31. Tobey K. L., Grant G. A. The nucleotide sequence of the serA gene of Escherichia coli and the amino acid sequence of the encoded protein, D-3-phosphoglycerate dehydrogenase. J Biol Chem. 1986 Sep 15;261(26):12179–12183. [PubMed] [Google Scholar]
  32. Turner J. M. Microbial metabolism of amino ketones. L-1-aminopropan-2-ol dehydrogenase and L-threonine dehydrogenase in Escherichia coli. Biochem J. 1967 Jul;104(1):112–121. doi: 10.1042/bj1040112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. UMBARGER H. E., UMBARGER M. A., SIU P. M. BIOSYNTHESIS OF SERINE IN ESCHERICHIA COLI AND SALMONELLA TYPHIMURIUM. J Bacteriol. 1963 Jun;85:1431–1439. doi: 10.1128/jb.85.6.1431-1439.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. URATA G., GRANICK S. Biosynthesis of alpha-aminoketones and the metabolism of aminoacetone. J Biol Chem. 1963 Feb;238:811–820. [PubMed] [Google Scholar]
  35. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  36. Wong H. C., Lessie T. G. Hydroxy amino acid metabolism in Pseudomonas cepacia: role of L-serine deaminase in dissimilation of serine, glycine, and threonine. J Bacteriol. 1979 Oct;140(1):240–245. doi: 10.1128/jb.140.1.240-245.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yang H. L., Ivashkiv L., Chen H. Z., Zubay G., Cashel M. Cell-free coupled transcription-translation system for investigation of linear DNA segments. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7029–7033. doi: 10.1073/pnas.77.12.7029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Zubay G. In vitro synthesis of protein in microbial systems. Annu Rev Genet. 1973;7:267–287. doi: 10.1146/annurev.ge.07.120173.001411. [DOI] [PubMed] [Google Scholar]

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