Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Screening of Growth- or Development-related Genes by Using Genomic Library with Inducible Promoter in Aspergillus nidulans

  • Lee Bang-Yong (Department of Biology, Graduate School of Wonkwang University) ;
  • Han Sang-Yong (Department of Biology, Graduate School of Wonkwang University) ;
  • Choi Han Gil (Department of Biology, Graduate School of Wonkwang University) ;
  • Kim Jee Hyun (Institute of Basic Natural Science, Wonkwang University) ;
  • Han Kap-Hoon (Research Center for Biomedicinal Resources, Pai Chai University) ;
  • Han Dong-Min (Institute of Basic Natural Science, Wonkwang University)
  • Published : 2005.12.01
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Abstract

Using the genomic library constructed at the downstream of the niiA promoter, which induces the over-expression of an inserted DNA fragment, we have attempted to screen the genes affecting growth or development by over-expression. The wild-type strain was transformed using the AMA-niiA(p) library and cultured on 1.2 M sorbitol media, in which asexual sporulation is induced, but sexual development is repressed. Over 100,000 strains transformed to $pyrG^+$ were analyzed with regard to any changes in phenotype. Consequently, seven strains were isolated for further analyses. These strains were designated NOT [niiA(p) over-expression transformants] stains. Four of the strains were of the inducible type, and the remaining strains were of the multi-copy suppression type. Two of the inducible-type strains, NOT 1 and NOT40, harbored genes which had been inserted in reverse direction, suggesting that the mutant phenotypes had been derived from an excess amount of anti-sense mRNA. Domain analyses of the deduced polypeptides from the DNA fragments rescued from the transformants revealed that NOT1, NOT40 and NOT6 harbored a LisH motif, a forkhead domain, and a $Zn(II)_2Cys_6$ binuclear zinc cluster, respectively.

Keywords

References

  1. Adams, T.H., J.K. Wieser, and J.H. Yu. 1998. Asexual sporulation in Aspergillus nidulans. Microbiol. Mol. Biol. Rev. 62, 35-54
  2. Carlsson, P. and M. Mahlapuu. 2002. Forkhead transcription factors: key players in development and metabolism. Dev. Biol. 250, 1-23 https://doi.org/10.1006/dbio.2002.0780
  3. Cottrell, T.R. and T.L. Doering. 2003. Silence of the strands: RNA interference in eukaryotic pathogens. Trends Microbiol. 11, 37-43 https://doi.org/10.1016/S0966-842X(02)00004-5
  4. Emes, R.D. and C.P. Ponting. 2001. A new sequence motif linking lissencephaly, Treacher Collins and oral-facial-digital type 1 syndromes, microtubule dynamics and cell migration. Hum. Mol. Gen. 10, 2813-2820 https://doi.org/10.1093/hmg/10.24.2813
  5. Han, D.M., Y.J. Han, Y.H. Lee, K.Y. Jahng, S.H. Jahng, and K.S. Chae. 1990. Inhibitory conditions of asexual development and their application for the screening mutants defective in sexual development. Kor. J. Mycol. 18, 225-232
  6. Han, K.-H., D.-B. Lee, J.-H. Kim, M.-S. Kim, K.-Y. Han, Y.-S. Park, W.-S. Kim, H.-B. Kim, and D.-M. Han. 2003. Environmental factors affecting sexual development of Aspergillus nidulans. J. Microbiol. 41, 34-40
  7. Han, K.-H. and R.A. Prade. 2002. Osmotic stress coupled maintenance of polar growth in Aspergillus nidulans. Mol. Microbiol. 43, 1065-1078 https://doi.org/10.1046/j.1365-2958.2002.02774.x
  8. Kafer, E. 1977. Meiotic and mitotic recombination in Aspergillus and its chromosomal aberrations. Adv. Genet. 19, 33-131 https://doi.org/10.1016/S0065-2660(08)60245-X
  9. Lee, S.B. and J.W. Taylor. 1990. Isolation of DNA from fungal mycelia and single spores, p. 282-287. In M.A. Innis, D.H. Gelfand, J.J. Sninsky and T.J. White (eds.), PCR Protocol; a Guide to Methods and Application, Academic Press, San Diego, California
  10. Masloff, S., S. Jacobsen, S. Poggeler, and U. Kuck. 2002. Functional analysis of the C6 zinc finger gene pro1 involved in fungal sexual development. Fungal Genet. Biol. 36, 107-116 https://doi.org/10.1016/S1087-1845(02)00010-5
  11. Mooney, J.L. and L.N. Yager. 1990. Light is required for conidiation in Aspergillus nidulans. Genes Dev. 4, 1473-1482 https://doi.org/10.1101/gad.4.9.1473
  12. Nakayashiki, H., S. Hanada, B.Q. Nguyen, N. Kadotani, Y. Tosa, and S. Mayama. 2005. RNA silencing as a tool for exploring gene function in ascomycete fungi. Fungal Genet. Biol. 42, 275-283 https://doi.org/10.1016/j.fgb.2005.01.002
  13. Osherov, N., J. Mathew, and G.S. May. 2000. Polarity-defective mutants of Aspergillus nidulans. Fungal Genet. Biol. 31, 181-188 https://doi.org/10.1006/fgbi.2000.1236
  14. Osherov, N. and G. May. 2000. Conidial germination in Aspergillus nidulans requires RAS signaling and protein synthesis. Genetics. 155, 647-656
  15. Pontecorvo, G., J.A. Roper, L.M. Hemmons, K.D. MacDonald, and A.W.J. Bufton. 1953. The genetics of Aspergillus nidulans. Adv. Genet. 5, 141-238 https://doi.org/10.1016/S0065-2660(08)60408-3
  16. Ray, A., S. Macwana, P. Ayoubi, L.T. Hall, R. Prade, and A.J. Mort. 2004. Negative subtraction hybridization: an efficient method to isolate large numbers of condition-specific cDNAs. BMC Genomics. 5, 22
  17. Sambrook, J.E., E.F. Fritsch, and T. Maniatis. 1987. Molecular cloning: A laboratory manual, 2nd ed. Cold Spring Harbor Laboratory Press, New York
  18. Schmitt, E.K., B. Hoff, and U. Kuck. 2004. AcFKH1, a novel member of the forkhead family, associates with the RFX transcription factor CPCR1 in the cephalosporin C-producing fungus Acremonium chrysogenum. Gene. 342, 269-281 https://doi.org/10.1016/j.gene.2004.08.002
  19. Shapira, M., E. Segal, and D. Botstein. 2004. Disruption of yeast forkhead-associated cell cycle transcription by oxidative stress. Mol. Biol. Cell. 15, 5659-5669 https://doi.org/10.1091/mbc.E04-04-0340
  20. Sims, A.H., G.D. Robson, D.C. Hoyle, S.G. Oliver, G. Turner, P.A. Prade, H.H. Russell, N.S. Dunn-Coleman, and M.E. Gent. 2004. Use of expressed sequence tag analysis and cDNA microarrays of the filamentous fungus Aspergillus nidulans. Fungal Genet. Biol. 41, 199-212 https://doi.org/10.1016/j.fgb.2003.11.005
  21. Szilagyi, Z., G. Batta, K. Enczi, and M. Sipiczki. 2005. Characterisation of two novel fork-head gene homologues of Schizosac-charomyces pombe: their involvement in cell cycle and sexual differentiation. Gene. 348, 101-109 https://doi.org/10.1016/j.gene.2004.12.043
  22. Timberlake, W.E. 1990. Molecular genetics of Aspergillus development. Annu. Rev. Genet. 24, 5-36 https://doi.org/10.1146/annurev.ge.24.120190.000253
  23. Vienken, K., M. Scherer, and R. Fischer. 2005. The Zn(II)2Cys6 putative Aspergillus nidulans transcription factor repressor of sexual development inhibits sexual development under lowcarbon conditions and in submersed culture. Genetics. 169, 619-630 https://doi.org/10.1534/genetics.104.030767
  24. Yelton, M.M., J.E. Hamer, and W.E. Timberlake. 1984. Transformation of Aspergillus nidulans by using a trpC plasmid. Proc. Natl. Acad. Sci. USA. 81, 1470-1474
  25. Yu, J.H., R.A. Butchko, M. Fernandes, N.P. Keller, T.J. Leonard, and T.H. Adams. 1996. Conservation of structure and function of the aflatoxin regulatory gene aflR from Aspergillus nidulans and A. flavus. Curr. Genet. 29, 549-555 https://doi.org/10.1007/BF02426959