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Formation of hybrid anthranilate synthetase in vitro from components of Aspergillus and Neurospora

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Abstract

Glutamine-dependent anthranilate synthetase was produced in vitro by mixing the extracts of a trypA and a trypC mutant of Aspergillus nidulans. Neither mutant alone possessed this activity. The enzyme formed in the mixture had the properties of the wild-type anthranilate synthetase which, together with N-(5′-phosphoribosyl) anthranilate (PRA) isomerase and indole 3-glycerol phosphate (InGP) synthetase, is found in a 10S multienzyme complex. Extracts of the trypA69 mutant contained a 6.5S protein as the active component—presumably the trypC + product—which in addition showed PRA isomerase and InGP synthetase activity. Extracts of the trypC801 mutant lacked all three enzyme activities but contained a 4.5S component—the trypA + gene product—which in vitro showed ammonia-dependent anthranilate synthetase activity. These mutants are analogous in their properties to certain tryp-2 and tryp-1 mutants of Neurospora. When complementary extracts of the two genera were mixed (Aspergillus trypA with Neurospora tryp-1 or Aspergillus trypC with Neurospora tryp-2), a “hybrid” glutamine-dependent anthranilate synthetase was obtained which showed less than half the activity produced in homologous combinations.

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Author notes

  1. J. A. DeMoss

    Present address: Biochemistry and Molecular Biology Group, University of Texas Medical School at Houston, Houston, Texas

Authors and Affiliations

  1. Department of Biology, McGill University, Montreal, Quebec, Canada

    Etta Käfer

  2. Department of Biology, San Diego University of California, La Jolla, California

    J. A. DeMoss

Authors
  1. Etta Käfer
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  2. J. A. DeMoss
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Additional information

This study was supported by Grant GB 22655 from the National Science Foundation to J.A.DeM.

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Käfer, E., DeMoss, J.A. Formation of hybrid anthranilate synthetase in vitro from components of Aspergillus and Neurospora . Biochem Genet 9, 203–211 (1973). https://doi.org/10.1007/BF00487451

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  • DOI: https://doi.org/10.1007/BF00487451

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