Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1974 Feb;117(2):502–508. doi: 10.1128/jb.117.2.502-508.1974

Localization of Two Functions of the Phosphoribosyl Anthranilate Transferase of Escherichia coli to Distinct Regions of the Polypeptide Chain

Ethel Noland Jackson a,1, Charles Yanofsky a
PMCID: PMC285540  PMID: 4590474

Abstract

The trpD gene specifies a polypeptide which has both glutamine amidotransferase and phosphoribosyl anthranilate (PRA) transferase activities. Deletions fusing segments of trpD to the gene preceding it in the operon, trpE, were selected in strains carrying various trpD point mutations. The selection procedure required both that a deletion enter trpE and that it restore the PRA transferase activity which the parent trpD point mutant lacked. Deletion mutants were found which had PRA transferase activity although the first third of trpD was deleted. The existence of the mutants proves that a terminal segment of trpD is sufficient to specify a polypeptide having PRA transferase activity. The location of the deletion end points on the genetic map of trpD defines the extent of the trpD segment required for PRA transferase activity. This segment did not overlap the initial region of trpD required to specify the glutamine amidotransferase function of the trpD polypeptide. These results support the hypothesis (M. Grieshaber and R. Bauerle, 1972; H. Zalkin and L. H. Hwang, 1971) that the bifunctional trpD polypeptide might have evolved by fusion of a gene specifying a glutamine amidotransferase with a gene directing PRA transferase synthesis.

Full text

PDF
502

Selected References

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

  1. Gibson F., Pittard J., Reich E. Ammonium ions as the source of nitrogen for tryptophan biosynthesis in whole cells of Escherichia coli. Biochim Biophys Acta. 1967 Apr 25;136(3):573–576. doi: 10.1016/0304-4165(67)90020-7. [DOI] [PubMed] [Google Scholar]
  2. Hwang L. H., Zalkin H. Multiple forms of anthranilate synthetase-anthranilate 5-phosphoribosylpyrophosphate phosphoribosyltransferase from Salmonella typhimurium. J Biol Chem. 1971 Apr 25;246(8):2338–2345. [PubMed] [Google Scholar]
  3. Ito J., Cox E. C., Yanofsky C. Anthranilate synthetase, an enzyme specified by the tryptophan operon of Escherichia coli: purification and characterization of component I. J Bacteriol. 1969 Feb;97(2):725–733. doi: 10.1128/jb.97.2.725-733.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ito J., Yanofsky C. Anthranilate synthetase, an enzyme specified by the tryptophan operon of Escherichia coli: Comparative studies on the complex and the subunits. J Bacteriol. 1969 Feb;97(2):734–742. doi: 10.1128/jb.97.2.734-742.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Kane J. F., Holmes W. M., Jensen R. A. Metabolic interlock. The dual function of a folate pathway gene as an extra-operonic gene of tryptophan biosynthesis. J Biol Chem. 1972 Mar 10;247(5):1587–1596. [PubMed] [Google Scholar]
  7. Kuhn J. C., Pabst M. J., Somerville R. L. Mutant strains of Escherichia coli K-12 exhibiting enhanced sensitivity to 5-methyltryptophan. J Bacteriol. 1972 Oct;112(1):93–101. doi: 10.1128/jb.112.1.93-101.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. La Scolea L. J., Jr, Balbinder E. Restoration of phosphoribosyl transferase activity by partially deleting the trpB gene in the tryptophan operon of Salmonella typhimurium. J Bacteriol. 1972 Nov;112(2):877–885. doi: 10.1128/jb.112.2.877-885.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Margolin P., Bauerle R. H. Determinants for regulation and initiation of expression of tryptophan genes. Cold Spring Harb Symp Quant Biol. 1966;31:311–320. doi: 10.1101/sqb.1966.031.01.041. [DOI] [PubMed] [Google Scholar]
  10. Nagano H., Zalkin H., Henderson E. J. The anthranilate synthetase-anthranilate-5-phosphorribosylpyrophosphate phosphoribosyltransferase aggregate. On the reaction mechanism of anthranilate synthetase from Salmonella typhimurium. J Biol Chem. 1970 Aug 10;245(15):3810–3820. [PubMed] [Google Scholar]
  11. Queener S. F., Gunsalus I. C. Anthranilate synthase enzyme system and complementation in Pseudomonas species. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1225–1232. doi: 10.1073/pnas.67.3.1225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Sarabhai A., Brenner S. A mutant which reinitiates the polypeptide chain after chain termination. J Mol Biol. 1967 Jul 14;27(1):145–162. doi: 10.1016/0022-2836(67)90357-9. [DOI] [PubMed] [Google Scholar]
  13. Shapiro J. A., Adhya S. L. The galactose operon of E. coli K-12. II. A deletion analysis of operon structure and polarity. Genetics. 1969 Jun;62(2):249–264. doi: 10.1093/genetics/62.2.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Spudich J. A., Horn V., Yanofsky C. On the production of deletions in the chromosome of Escherichia coli. J Mol Biol. 1970 Oct 14;53(1):49–67. doi: 10.1016/0022-2836(70)90045-8. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]

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

RESOURCES