Skip to main content
Genetics logoLink to Genetics
. 1995 Feb;139(2):537–547. doi: 10.1093/genetics/139.2.537

Identification of Developmental Regulatory Genes in Aspergillus Nidulans by Overexpression

J F Marhoul 1, T H Adams 1
PMCID: PMC1206365  PMID: 7713416

Abstract

Overexpression of several Aspergillus nidulans developmental regulatory genes has been shown to cause growth inhibition and development at inappropriate times. We set out to identify previously unknown developmental regulators by constructing a nutritionally inducible A. nidulans expression library containing small, random genomic DNA fragments inserted next to the alcA promoter [ alcA (p) ] in an A. nidulans transformation vector. Among 20,000 transformants containing random alcA (p) genomic DNA fusion constructs, we identified 66 distinct mutant strains in which alcA (p) induction resulted in growth inhibition as well as causing other detectable phenotypic changes. These growth inhibited mutants were divided into 52 FIG (Forced expression Inhibition of Growth) and 14 FAB (Forced expression Activation of brlA) mutants based on whether or not alcA (p) induction resulted in accumulation of mRNA for the developmental regulatory gene brlA. In four FAB mutants, alcA (p) induction not only activated brlA expression but also caused hyphae to differentiate into reduced conidiophores that produced viable spores from the tips as is observed after alcA (p) :: brlA induction. Sequence analyses of the DNA fragments under alcA (p) control in three of these four sporulating strains showed that in two cases developmental activation resulted from overexpression of previously uncharacterized genes, whereas in the third strain, the alcA (p) was fused to brlA. The potential uses for this strategy in identifying genes whose overexpression results in specific phenotypic changes like developmental induction are discussed.

Full Text

The Full Text of this article is available as a PDF (5.7 MB).

Selected References

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

  1. Adams T. H., Boylan M. T., Timberlake W. E. brlA is necessary and sufficient to direct conidiophore development in Aspergillus nidulans. Cell. 1988 Jul 29;54(3):353–362. doi: 10.1016/0092-8674(88)90198-5. [DOI] [PubMed] [Google Scholar]
  2. Adams T. H., Timberlake W. E. Developmental repression of growth and gene expression in Aspergillus. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5405–5409. doi: 10.1073/pnas.87.14.5405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  4. Boylan M. T., Mirabito P. M., Willett C. E., Zimmerman C. R., Timberlake W. E. Isolation and physical characterization of three essential conidiation genes from Aspergillus nidulans. Mol Cell Biol. 1987 Sep;7(9):3113–3118. doi: 10.1128/mcb.7.9.3113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burke D., Gasdaska P., Hartwell L. Dominant effects of tubulin overexpression in Saccharomyces cerevisiae. Mol Cell Biol. 1989 Mar;9(3):1049–1059. doi: 10.1128/mcb.9.3.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clutterbuck A. J. A mutational analysis of conidial development in Aspergillus nidulans. Genetics. 1969 Oct;63(2):317–327. doi: 10.1093/genetics/63.2.317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dietzel C., Kurjan J. The yeast SCG1 gene: a G alpha-like protein implicated in the a- and alpha-factor response pathway. Cell. 1987 Sep 25;50(7):1001–1010. doi: 10.1016/0092-8674(87)90166-8. [DOI] [PubMed] [Google Scholar]
  8. Gwynne D. I., Buxton F. P., Sibley S., Davies R. W., Lockington R. A., Scazzocchio C., Sealy-Lewis H. M. Comparison of the cis-acting control regions of two coordinately controlled genes involved in ethanol utilization in Aspergillus nidulans. Gene. 1987;51(2-3):205–216. doi: 10.1016/0378-1119(87)90309-x. [DOI] [PubMed] [Google Scholar]
  9. Haarer B. K., Pringle J. R. Immunofluorescence localization of the Saccharomyces cerevisiae CDC12 gene product to the vicinity of the 10-nm filaments in the mother-bud neck. Mol Cell Biol. 1987 Oct;7(10):3678–3687. doi: 10.1128/mcb.7.10.3678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Herskowitz I. Functional inactivation of genes by dominant negative mutations. Nature. 1987 Sep 17;329(6136):219–222. doi: 10.1038/329219a0. [DOI] [PubMed] [Google Scholar]
  11. Hurt E. C. A novel nucleoskeletal-like protein located at the nuclear periphery is required for the life cycle of Saccharomyces cerevisiae. EMBO J. 1988 Dec 20;7(13):4323–4334. doi: 10.1002/j.1460-2075.1988.tb03331.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Käfer E. Meiotic and mitotic recombination in Aspergillus and its chromosomal aberrations. Adv Genet. 1977;19:33–131. doi: 10.1016/s0065-2660(08)60245-x. [DOI] [PubMed] [Google Scholar]
  13. Lee B. N., Adams T. H. Overexpression of flbA, an early regulator of Aspergillus asexual sporulation, leads to activation of brlA and premature initiation of development. Mol Microbiol. 1994 Oct;14(2):323–334. doi: 10.1111/j.1365-2958.1994.tb01293.x. [DOI] [PubMed] [Google Scholar]
  14. Liu H., Krizek J., Bretscher A. Construction of a GAL1-regulated yeast cDNA expression library and its application to the identification of genes whose overexpression causes lethality in yeast. Genetics. 1992 Nov;132(3):665–673. doi: 10.1093/genetics/132.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mirabito P. M., Adams T. H., Timberlake W. E. Interactions of three sequentially expressed genes control temporal and spatial specificity in Aspergillus development. Cell. 1989 Jun 2;57(5):859–868. doi: 10.1016/0092-8674(89)90800-3. [DOI] [PubMed] [Google Scholar]
  16. Osmani S. A., Engle D. B., Doonan J. H., Morris N. R. Spindle formation and chromatin condensation in cells blocked at interphase by mutation of a negative cell cycle control gene. Cell. 1988 Jan 29;52(2):241–251. doi: 10.1016/0092-8674(88)90513-2. [DOI] [PubMed] [Google Scholar]
  17. Osmani S. A., Pu R. T., Morris N. R. Mitotic induction and maintenance by overexpression of a G2-specific gene that encodes a potential protein kinase. Cell. 1988 Apr 22;53(2):237–244. doi: 10.1016/0092-8674(88)90385-6. [DOI] [PubMed] [Google Scholar]
  18. PONTECORVO G., ROPER J. A., HEMMONS L. M., MACDONALD K. D., BUFTON A. W. J. The genetics of Aspergillus nidulans. Adv Genet. 1953;5:141–238. doi: 10.1016/s0065-2660(08)60408-3. [DOI] [PubMed] [Google Scholar]
  19. Rose M. D., Fink G. R. KAR1, a gene required for function of both intranuclear and extranuclear microtubules in yeast. Cell. 1987 Mar 27;48(6):1047–1060. doi: 10.1016/0092-8674(87)90712-4. [DOI] [PubMed] [Google Scholar]
  20. Stringer M. A., Dean R. A., Sewall T. C., Timberlake W. E. Rodletless, a new Aspergillus developmental mutant induced by directed gene inactivation. Genes Dev. 1991 Jul;5(7):1161–1171. doi: 10.1101/gad.5.7.1161. [DOI] [PubMed] [Google Scholar]
  21. Timberlake W. E., Marshall M. A. Genetic engineering of filamentous fungi. Science. 1989 Jun 16;244(4910):1313–1317. doi: 10.1126/science.2525275. [DOI] [PubMed] [Google Scholar]
  22. Timberlake W. E. Molecular genetics of Aspergillus development. Annu Rev Genet. 1990;24:5–36. doi: 10.1146/annurev.ge.24.120190.000253. [DOI] [PubMed] [Google Scholar]
  23. Upshall A., Gilbert T., Saari G., O'Hara P. J., Weglenski P., Berse B., Miller K., Timberlake W. E. Molecular analysis of the argB gene of Aspergillus nidulans. Mol Gen Genet. 1986 Aug;204(2):349–354. doi: 10.1007/BF00425521. [DOI] [PubMed] [Google Scholar]
  24. Yager L. N., Kurtz M. B., Champe S. P. Temperature-shift analysis of conidial development in Aspergillus nidulans. Dev Biol. 1982 Sep;93(1):92–103. doi: 10.1016/0012-1606(82)90242-1. [DOI] [PubMed] [Google Scholar]
  25. Yelton M. M., Hamer J. E., Timberlake W. E. Transformation of Aspergillus nidulans by using a trpC plasmid. Proc Natl Acad Sci U S A. 1984 Mar;81(5):1470–1474. doi: 10.1073/pnas.81.5.1470. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES