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. 1976 May;126(2):668–678. doi: 10.1128/jb.126.2.668-678.1976

Naturally occurring sites within the Shigella dysenteriae tryptophan operon severely limit tryptophan biosynthesis.

M D Manson, C Yanofsky
PMCID: PMC233200  PMID: 770449

Abstract

We investigated the structural, functional, and regulatory properties of the Shigella dysenteriae tryptophan (trp.) operon in transduction hybrids in which the cysB-trp-region of Escherichia coli is replaced by the corresponding region from S. dysenteriae. Tryptophan biosynthesis was largely blocked in the hybrids, although the order of the structural genes was identical with that of E. coli. Nutritional tests and enzyme assays revealed that the hybrids produced a defective anthranilate synthetase (ASase). Deletion mapping identified two distinct sites in trpE, each of which was partially responsible for the instability and low activity of ASase. We also discovered a pleiotropic site trpP (S) that maps outside the structural gene region and is closely linked to the S. dysenteriae trp operator. trpP (S) reduced the rate of trp messenger ribonucleic acid synthesis, and consequently trp enzyme levels, 10-fold relative to wild-type E. coli. In recombinants in which the structural genes of E coli were under the control of the S. dysenteriae promoter, enzyme levels were also reduced 10-fold. In some fast-growing revertants of the original hybrids, the rates of trp messenger ribonucleic acid synthesis and levels of tryptophan synthetase were restored to values characteristic of wild-type E.coli. Thus, the Trp auxotrophy associated with the S dysenteriae trp operon derives from the combination of a defective ASase and decreased expression of the entire operon imposed by trpP (S).

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Selected References

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  1. Bertrand K., Korn L., Lee F., Platt T., Squires C. L., Squires C., Yanofsky C. New features of the regulation of the tryptophan operon. Science. 1975 Jul 4;189(4196):22–26. doi: 10.1126/science.1094538. [DOI] [PubMed] [Google Scholar]
  2. Brenner D. J., Fanning G. R., Skerman F. J., Falkow S. Polynucleotide sequence divergence among strains of Escherichia coli and closely related organisms. J Bacteriol. 1972 Mar;109(3):953–965. doi: 10.1128/jb.109.3.953-965.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bronson M. J., Squires C., Yanofsky C. Nucleotide sequences from tryptophan messenger RNA of Escherichia coli: the sequence corresponding to the amino-terminal region of the first polypeptide specified by the operon. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2335–2339. doi: 10.1073/pnas.70.8.2335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bronson M. J., Yanofsky C. Letter to the editor: Characterization of mutations in the tryptophan operon of Escherichia coli by RNA nucleotide sequencing. J Mol Biol. 1974 Oct 5;88(4):913–915. doi: 10.1016/0022-2836(74)90407-0. [DOI] [PubMed] [Google Scholar]
  5. De Crombrugghe B., Adhya S., Gottesman M., Pastan I. Effect of Rho on transcription of bacterial operons. Nat New Biol. 1973 Feb 28;241(113):260–264. doi: 10.1038/newbio241260a0. [DOI] [PubMed] [Google Scholar]
  6. Deeb S. S., Okamoto K., Hall B. D. Isolation and characterization of non-defective transducing elements of bacteriophage phi-80. Virology. 1967 Feb;31(2):289–295. doi: 10.1016/0042-6822(67)90173-0. [DOI] [PubMed] [Google Scholar]
  7. Denney R. M., Yanofsky C. Detection of tryptophan messenger RNA in several bacterial species and examination of the properties of heterologous DNA-RNA hybrids. J Mol Biol. 1972 Mar 14;64(2):319–339. doi: 10.1016/0022-2836(72)90501-3. [DOI] [PubMed] [Google Scholar]
  8. EISENSTEIN R. B., YANOFSKY C. Tryptophan synthetase levels in Escherichia coli, Shigella dysenteriae, and transduction hybrids. J Bacteriol. 1962 Jan;83:193–204. doi: 10.1128/jb.83.1.193-204.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Englesberg E., Ingraham L. MEIOTROPHIC MUTANTS OF Pasteurella Pestis AND THEIR USE IN THE ELUCIDATION OF NUTRITIONAL REQUIREMENTS. Proc Natl Acad Sci U S A. 1957 May 15;43(5):369–372. doi: 10.1073/pnas.43.5.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. FRANKLIN N. C., LURIA S. E. Transduction by bacteriophage P-1 and the properties of the lac genetic region in E. coli and S. dysenteriae. Virology. 1961 Nov;15:299–311. doi: 10.1016/0042-6822(61)90362-2. [DOI] [PubMed] [Google Scholar]
  11. Fiandt M., Szybalski W., Malamy M. H. Polar mutations in lac, gal and phage lambda consist of a few IS-DNA sequences inserted with either orientation. Mol Gen Genet. 1972;119(3):223–231. doi: 10.1007/BF00333860. [DOI] [PubMed] [Google Scholar]
  12. Hegeman G. D., Rosenberg S. L. The evolution of bacterial enzyme systems. Annu Rev Microbiol. 1970;24:429–462. doi: 10.1146/annurev.mi.24.100170.002241. [DOI] [PubMed] [Google Scholar]
  13. Hiraga S., Yanofsky C. Hyper-labile messenger RNA in polar mutants of the tryptophan operon of Escherichia coli. J Mol Biol. 1972 Dec 14;72(1):103–110. doi: 10.1016/0022-2836(72)90072-1. [DOI] [PubMed] [Google Scholar]
  14. Hiraga S., Yanofsky C. Normal repression in a deletion mutant lacking almost the entire operator-proximal gene of the tryptophan operon of E. coli. Nat New Biol. 1972 May 10;237(71):47–49. doi: 10.1038/newbio237047a0. [DOI] [PubMed] [Google Scholar]
  15. Hirsch H. J., Saedler H., Starlinger P. Insertion mutations in the control region of the galactose operon of E. coli. II. Physical characterization of the mutations. Mol Gen Genet. 1972;115(3):266–276. doi: 10.1007/BF00268890. [DOI] [PubMed] [Google Scholar]
  16. Hirsch H. J., Starlinger P., Brachet P. Two kinds of insertions in bacterial genes. Mol Gen Genet. 1972;119(3):191–206. doi: 10.1007/BF00333858. [DOI] [PubMed] [Google Scholar]
  17. Ito J., Crawford I. P. Regulation of the enzymes of the tryptophan pathway in Escherichia coli. Genetics. 1965 Dec;52(6):1303–1316. doi: 10.1093/genetics/52.6.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jackson E. N., Yanofsky C. Internal deletions in the tryptophan operon of Escherichia coli. J Mol Biol. 1972 Nov 14;71(2):149–161. doi: 10.1016/0022-2836(72)90343-9. [DOI] [PubMed] [Google Scholar]
  19. Jackson E. N., Yanofsky C. Localization of two functions of the phosphoribosyl anthranilate transferase of Escherichia coli to distinct regions of the polypeptide chain. J Bacteriol. 1974 Feb;117(2):502–508. doi: 10.1128/jb.117.2.502-508.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jackson E. N., Yanofsky C. Thr region between the operator and first structural gene of the tryptophan operon of Escherichia coli may have a regulatory function. J Mol Biol. 1973 May 5;76(1):89–101. doi: 10.1016/0022-2836(73)90082-x. [DOI] [PubMed] [Google Scholar]
  21. Koch A. L. Enzyme evolution. I. The importance of untranslatable intermediates. Genetics. 1972 Oct;72(2):297–316. doi: 10.1093/genetics/72.2.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Koch A. L. The adaptive responses of Escherichia coli to a feast and famine existence. Adv Microb Physiol. 1971;6:147–217. doi: 10.1016/s0065-2911(08)60069-7. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. LURIA S. E., BURROUS J. W. Hybridization between Escherichia coli and Shigella. J Bacteriol. 1957 Oct;74(4):461–476. doi: 10.1128/jb.74.4.461-476.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Li S. L., Yanofsky C. Amino acid sequences of fifty residues from the amino termini of the tryptophan synthetase chains of several enterobacteria. J Biol Chem. 1972 Feb 25;247(4):1031–1037. [PubMed] [Google Scholar]
  27. MCCARTHY B. J., BOLTON E. T. An approach to the measurement of genetic relatedness among organisms. Proc Natl Acad Sci U S A. 1963 Jul;50:156–164. doi: 10.1073/pnas.50.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Manson M. D., Yanofsky C. Tryptophan operon regulation in interspecific hybrids of enteric bacteria. J Bacteriol. 1976 May;126(2):679–689. doi: 10.1128/jb.126.2.679-689.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mojica-A T., Middleton R. B. Fertility of Salmonella typhimurium crosses with Escherichia coli. J Bacteriol. 1971 Dec;108(3):1161–1167. doi: 10.1128/jb.108.3.1161-1167.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Morishita T., Fukada T., Shirota M., Yura T. Genetic basis of nutritional requirements in Lactobacillus casei. J Bacteriol. 1974 Dec;120(3):1078–1084. doi: 10.1128/jb.120.3.1078-1084.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Morse D. E., Yanofsky C. A transcription-initiating mutation within a structural gene of the tryptophan operon. J Mol Biol. 1969 May 14;41(3):317–328. doi: 10.1016/0022-2836(69)90278-2. [DOI] [PubMed] [Google Scholar]
  32. Morse D. E., Yanofsky C. Polarity and the degradation of mRNA. Nature. 1969 Oct 25;224(5217):329–331. doi: 10.1038/224329a0. [DOI] [PubMed] [Google Scholar]
  33. Murgola E. J., Yanofsky C. Selection for new amino acids at position 211 of the tryptophan synthetase alpha chain of Escherichia coli. J Mol Biol. 1974 Jul 15;86(4):775–784. doi: 10.1016/0022-2836(74)90353-2. [DOI] [PubMed] [Google Scholar]
  34. Saedler H., Besemer J., Kemper B., Rosenwirth B., Starlinger P. Insertion mutations in the control region of the Gal operon of E. coli. I. Biological characterization of the mutations. Mol Gen Genet. 1972;115(3):258–265. doi: 10.1007/BF00268889. [DOI] [PubMed] [Google Scholar]
  35. Sarkar S. Properties and regulation of the beta-D-galactosidase in Shigella dysenteriae and in Escherichia coli-Shigella dysenteriae hybrids. J Bacteriol. 1966 Apr;91(4):1477–1488. doi: 10.1128/jb.91.4.1477-1488.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. YANOFSKY C., LENNOX E. S. Transduction and recombination study of linkage relationships among the genes controlling tryptophan synthesis in Escherichia coli. Virology. 1959 Aug;8:425–447. doi: 10.1016/0042-6822(59)90046-7. [DOI] [PubMed] [Google Scholar]
  38. Yanofsky C., Horn V., Bonner M., Stasiowski S. Polarity and enzyme functions in mutants of the first three genes of the tryptophan operon of Escherichia coli. Genetics. 1971 Dec;69(4):409–433. doi: 10.1093/genetics/69.4.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Yanofsky C., Ito J. Nonsense codons and polarity in the tryptophan operon. J Mol Biol. 1966 Nov 14;21(2):313–334. doi: 10.1016/0022-2836(66)90102-1. [DOI] [PubMed] [Google Scholar]
  40. Zalkin H., Kling D. Anthranilate synthetase. Purification and properties of component I from Salmonella typhimurium. Biochemistry. 1968 Oct;7(10):3566–3573. doi: 10.1021/bi00850a034. [DOI] [PubMed] [Google Scholar]
  41. Zamenhof S., Eichhorn H. H. Study of microbial evolution through loss of biosynthetic functions: establishment of "defective" mutants. Nature. 1967 Nov 4;216(5114):456–458. doi: 10.1038/216456a0. [DOI] [PubMed] [Google Scholar]

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