Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Jun;84(11):3753–3757. doi: 10.1073/pnas.84.11.3753

Complementation of areA- regulatory gene mutations of Aspergillus nidulans by the heterologous regulatory gene nit-2 of Neurospora crassa.

M A Davis, M J Hynes
PMCID: PMC304954  PMID: 2954160

Abstract

Loss-of-function mutations in the regulatory gene areA of Aspergillus nidulans prevent the utilization of a wide variety of nitrogen sources. The phenotypes of nit-2 mutants of Neurospora crassa suggest that this gene may be analogous to the areA gene. Transformation has been used to introduce a plasmid containing the nit-2 gene into A. nidulans. The nit-2 gene of Neurospora complemented mutations in the areA gene, restoring the ability to use a variety of nitrogen sources. This indicated that the activator function of nit-2 and areA gene products was retained across these two fungal species. Southern blot analysis revealed both single-copy and multicopy integrations and, in at least one case, integration appeared to generate a nit-2 mutation. Integration of the transforming plasmid appeared to be by nonhomologous events at a number of different sites in the Aspergillus genome. The transformants were less sensitive to nitrogen-metabolite repression of extracellular protease activity and nitrate reductase (EC 1.6.6.3) than were wild-type A. nidulans. This indicated that nitrogen control was not completely normal in the nit-2 transformants.

Full text

PDF
3755

Images in this article

3755Image
on p.3755
3755Image
on p.3755
3756Image
on p.3756

Selected References

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

  1. Arst H. N., Jr A near terminal pericentric inversion leads to nitrogen metabolite derepression in Aspergillus nidulans. Mol Gen Genet. 1982;188(3):490–493. doi: 10.1007/BF00330054. [DOI] [PubMed] [Google Scholar]
  2. Arst H. N., Jr, Cove D. J. Nitrogen metabolite repression in Aspergillus nidulans. Mol Gen Genet. 1973 Nov 2;126(2):111–141. doi: 10.1007/BF00330988. [DOI] [PubMed] [Google Scholar]
  3. 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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  4. Caddick M. X., Arst H. N., Jr, Taylor L. H., Johnson R. I., Brownlee A. G. Cloning of the regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans. EMBO J. 1986 May;5(5):1087–1090. doi: 10.1002/j.1460-2075.1986.tb04326.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chada K., Magram J., Costantini F. An embryonic pattern of expression of a human fetal globin gene in transgenic mice. Nature. 1986 Feb 20;319(6055):685–689. doi: 10.1038/319685a0. [DOI] [PubMed] [Google Scholar]
  6. Cove D. J. The induction and repression of nitrate reductase in the fungus Aspergillus nidulans. Biochim Biophys Acta. 1966 Jan 11;113(1):51–56. doi: 10.1016/s0926-6593(66)80120-0. [DOI] [PubMed] [Google Scholar]
  7. DeBusk R. M., Ogilvie S. Regulation of amino acid utilization in Neurospora crassa: effect of nmr-1 and ms-5 mutations. J Bacteriol. 1984 Nov;160(2):656–661. doi: 10.1128/jb.160.2.656-661.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Durrens P., Green P. M., Arst H. N., Jr, Scazzocchio C. Heterologous insertion of transforming DNA and generation of new deletions associated with transformation in Aspergillus nidulans. Mol Gen Genet. 1986 Jun;203(3):544–549. doi: 10.1007/BF00422084. [DOI] [PubMed] [Google Scholar]
  9. Gurr S. J., Hawkins A. R., Drainas C., Kinghorn J. R. Isolation and identification of the Aspergillus nidulans gdhA gene encoding NADP-linked glutamate dehydrogenase. Curr Genet. 1986;10(10):761–766. doi: 10.1007/BF00405099. [DOI] [PubMed] [Google Scholar]
  10. Hynes M. J., Corrick C. M., King J. A. Isolation of genomic clones containing the amdS gene of Aspergillus nidulans and their use in the analysis of structural and regulatory mutations. Mol Cell Biol. 1983 Aug;3(8):1430–1439. doi: 10.1128/mcb.3.8.1430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hynes M. J., Pateman J. A. The genetic analysis of regulation of amidase synthesis in Aspergillus nidulans. I. Mutants able to utilize acrylamide. Mol Gen Genet. 1970;108(2):97–106. doi: 10.1007/BF02430516. [DOI] [PubMed] [Google Scholar]
  12. Hynes M. J., Pateman J. A. The genetic analysis of regulation of amidase synthesis in Aspergillus nidulans. II. Mutants resistant to fluoroacetamide. Mol Gen Genet. 1970;108(2):107–116. doi: 10.1007/BF02430517. [DOI] [PubMed] [Google Scholar]
  13. Hynes M. J. Studies on the role of the areA gene in the regulation of nitrogen catabolism in Aspergillus nidulans. Aust J Biol Sci. 1975 Jun;28(3):301–313. doi: 10.1071/bi9750301. [DOI] [PubMed] [Google Scholar]
  14. Lamppa G., Nagy F., Chua N. H. Light-regulated and organ-specific expression of a wheat Cab gene in transgenic tobacco. Nature. 1985 Aug 22;316(6030):750–752. doi: 10.1038/316750a0. [DOI] [PubMed] [Google Scholar]
  15. Lewis C. M., Fincham J. R. Regulation of nitrate reductase in the basidiomycete Ustilago maydis. J Bacteriol. 1970 Jul;103(1):55–61. doi: 10.1128/jb.103.1.55-61.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Marzluf G. A. Regulation of nitrogen metabolism and gene expression in fungi. Microbiol Rev. 1981 Sep;45(3):437–461. doi: 10.1128/mr.45.3.437-461.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Perrine K. G., Marzluf G. A. Amber nonsense mutations in regulatory and structural genes of the nitrogen control circuit of Neurospora crassa. Curr Genet. 1986;10(9):677–684. doi: 10.1007/BF00410916. [DOI] [PubMed] [Google Scholar]
  18. Spena A., Hain R., Ziervogel U., Saedler H., Schell J. Construction of a heat-inducible gene for plants. Demonstration of heat-inducible activity of the Drosophila hsp70 promoter in plants. EMBO J. 1985 Nov;4(11):2739–2743. doi: 10.1002/j.1460-2075.1985.tb03997.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Stewart V., Vollmer S. J. Molecular cloning of nit-2, a regulatory gene required for nitrogen metabolite repression in Neurospora crassa. Gene. 1986;46(2-3):291–295. doi: 10.1016/0378-1119(86)90414-2. [DOI] [PubMed] [Google Scholar]
  20. Tilburn J., Scazzocchio C., Taylor G. G., Zabicky-Zissman J. H., Lockington R. A., Davies R. W. Transformation by integration in Aspergillus nidulans. Gene. 1983 Dec;26(2-3):205–221. doi: 10.1016/0378-1119(83)90191-9. [DOI] [PubMed] [Google Scholar]
  21. Wiame J. M., Grenson M., Arst H. N., Jr Nitrogen catabolite repression in yeasts and filamentous fungi. Adv Microb Physiol. 1985;26:1–88. doi: 10.1016/s0065-2911(08)60394-x. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES