Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Isolation and Characterization of the gtfA Gene Encoding GAL4-Like Transcription Factor in Aspergillus nidulans

Aspergillus nidulans에서 GAL4 유사 전사인자를 암호화하는 gtfA 유전자의 분리 및 분석

  • Received : 2013.03.08
  • Accepted : 2013.03.25
  • Published : 2013.03.31
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Abstract

A GAL4 type transcription factor gene (formally annotated as AN3912) locating downstream of sndA (AN3911) was characterized. The putative transcription factor carries both Zn(II)2Cys6 binuclear cluster DNA-binding domain and transcription activator domain. The gene named gtfA (gal4 type transcription factor) had an open reading frame which consisted of 762 amino acids and was disrupted by three introns. The deletion mutant produced reduced amount of conidia but increased amount of fruiting bodies, suggesting that the GtfA make function in decision of asexual preferential to sexual development. The forced over expression of gtfA caused the retardation of fruiting body formation on high glucose concentration. The transcript level of gtfA was kept constant through the life cycle except late vegetative stage and early sexual development stage during which slight increase was found. The expression of gtfA was not significantly affected by sexual or asexual development regulators, such as VeA, NsdD or FluG, FadA, and SfaD. The GtfA repressed the nsdC transcription, which suggested that GftA control sexual development negatively via negative regulation of nsdC expression.

sndA 유전자(AN3911) 하위에 위치하고 있는 GAL4형 전사인자를 암호화하는 유전자(AN3912)에 대해 분석하였다. 이 전사인자는 Zn(II)2Cys6 binuclear cluster DNA-binding domain 과 전사활성부위를 모두 가지고 있는 전형적 진균 특이 전사인자로서 해당 유전자는 gtfA (gal4 type transcription factor)라 명명되었다. 이 유전자의 ORF는 762개의 아미노산으로 구성되어 있고 3개의 intron이 그 안에 존재하였다. gtfA 유전자의 결실돌연변이는 무성포자생성이 감소하고 대신 유성생식기관이 증가하는 형질을 보여주었다. 과다발현균주는 높은 포도당 농도가 주어지면 유성생식이 늦어지는 형질을 나타내었다. gtfA 유전자는 영양생장 후반부와 유성분화 초기단계에서 높은 발현량을 보이고 전생활사를 통해 비교적 일정하게 발현되었다. gtfA의 전사는 일부 유성(NsdD, VeA) 및 무성(FluG, FadA, SfaD) 분화 조절자들에 의해 크게 영향을 받지 않았다. 반면 GtfA는 nsdC 유전자의 발현을 억제하는 것으로 나타났다. 종합하여 볼 때, GtfA는 분화 결정시기에 nsdC의 발현을 억제함으로써 유성분화보다는 무성분화로 유도되도록 조절 할 것으로 추정된다.

Keywords

References

  1. Adams, T.H. and Yu, J.H. 1998. Coordinate control of secondary metabolite production and asexual sporulation in Aspergillus nidulans. Curr. Opin. Microbiol. 1, 674-677. https://doi.org/10.1016/S1369-5274(98)80114-8
  2. Andrianopoulos, A. and Timberlake, W.E. 1991. ATTS, a new and conserved DNA binding domain. Plant Cell. 3, 747-748. https://doi.org/10.1105/tpc.3.8.747
  3. Boylan, M.T., Mirabio, M., Willett, C.E., Zimmerman, C.R., and Timberlake, W.E. 1987. Isolation and physical characterization of three essential conidiation genes from Aspergillus nidulans. Mol. Cell. Biol. 7, 3113-3118. https://doi.org/10.1128/MCB.7.9.3113
  4. Burger, G., Strauss, J., Scazzocchio, C., and Lang, B.F. 1991. nirA, the pathway specific regulatory gene of nitrate assimilation in Aspergillus nidulans, encodes a putative GAL4-type zinc finger protein and contains four introns in highly conserved regions. Mol. Cell. Biol. 11, 5746-5755. https://doi.org/10.1128/MCB.11.11.5746
  5. Byram, O., Krappmann, S., Ni, M., Bok, J.W., Helmstaedt, K., Valerius, O., Braus-Stromeyer, S., Kwon, N.J., Keller, N.P., Yu, J.H., and et al. 2008. VelB/VeA/LaeA complex coordinates light signal with fungal development and secondary metabolism. Science 320, 1504-1506. https://doi.org/10.1126/science.1155888
  6. Cary, J.W., Harris-Coward, P.Y., Ehrlich, K.C., Mack, B.M., Kale, S.P., Larey, C., and Calvo, A.M. 2012. NsdC and NsdD affect Aspergillus flavus morphogenesis and aflatoxin production. Eukaryot. Cell. 11, 1104-1011. https://doi.org/10.1128/EC.00069-12
  7. Davis, M.A., Small, A.J., Kourambis, S., and Hynes, M.J. 1996. The tamA gene of Aspergillus nidulans contains a putative zinc cluster motif which is not required for gene function. J. Bacteriol. 178, 3406 -3409. https://doi.org/10.1128/jb.178.11.3406-3409.1996
  8. Gehring, W.J. 1992. The homeobox in perspective. Trends Biochem. Sci. 17, 277-280. https://doi.org/10.1016/0968-0004(92)90434-B
  9. Han, D.M., Han, Y.J., Lee, Y.H., Jahng, K.Y., Jahng, S.H., and Chae, K.S. 1990. Inhibitory conditions of asexual development and their application for the screening of mutants defective in sexual development. Kor. J. Mycol. 18, 225-232.
  10. Han, K.H., Han, K.Y., Yu, J.H., Chae, K.S., Jahng, K.Y., and Han, D.M. 2001. The nsdD gene encodes a putative GATA-type transcription factor necessary for sexual development of Aspergillus nidulans. Mol. Microbiol. 41, 299-309. https://doi.org/10.1046/j.1365-2958.2001.02472.x
  11. Harley, V.R., Jackson, D.I., Hextall, P.J., Hawkins, J.R., Berkovitz,G.D., Sockanathan, S., Lovell-Badge, R., and Goodfellow, P.N. 1992. DNA binding activity of recombinant SRY from normal males and XY females. Science 255, 453-456. https://doi.org/10.1126/science.1734522
  12. Harrison, S.C. 1991. A structural taxonomy of DNA-binding domains. Nature 353, 715-719. https://doi.org/10.1038/353715a0
  13. Kim, H.R., Chae, K.S., Han, K.H., and Han, D.M. 2009. The nsdC gene encoding a putative C2H2-type transcription factor is a key activator of sexual development in Aspergillus nidulans. Genetics 182, 771-783. https://doi.org/10.1534/genetics.109.101667
  14. Kim, H.S., Han, K.Y., Kim, K.J., Han, D.M., Jahng, K.Y., and Chae, K.S. 2002. The veA gene activates sexual development in Aspergillus nidulans. Fungal Genet. Biol. 37, 72-80. https://doi.org/10.1016/S1087-1845(02)00029-4
  15. Lafon, A., Seo, J.A., Han, K.H., Yu, J.H., and d'Enfert, C. 2005. The heterotrimeric G-protein GanB(${\alpha}$)-SfaD(${\beta}$)-GpgA(${\gamma}$) is a carbon source sensor involved in early cAMP-dependent germination in Aspergillus nidulans. Genetics 171, 71-80. https://doi.org/10.1534/genetics.105.040584
  16. Lee, B.N. and Adams, T.H. 1994. The Aspergillus nidulans fluG gene is required for production of an extracellular developmental signal. Genes Dev. 8, 641-651. https://doi.org/10.1101/gad.8.6.641
  17. Lee, D.B., Kim, L.H., Kim, J.P., Han, K.H., and Han, D.M. 2011. Isolation and characterization of self-fertile suppressors from the sterile nsdD deletion mutant of Aspergillus nidulans. J. Microbiol. 49, 1054-1057. https://doi.org/10.1007/s12275-011-1111-4
  18. Murre, C., Bain, G., van Dijk, M.A., Engel, I., Furnari, B.A., Massari, M.E., Matthews, J.R., Quong, M.W., Rivera, R.R., and Stuiver, M.H. 1994. Structure of helix-loop-helix proteins. Biochim. Biophys. Acta. 1218, 129-135. https://doi.org/10.1016/0167-4781(94)90001-9
  19. Punt, P.J., Strauss, J., Smit, R., Kinghorn, J.R., van den Hondel, C.A.M.J.J., and Scazzocchio, C. 1995. The intergenic region between the divergently transcribed niiA and niaD gene of Aspergillus nidulans contains multiple NirA binding sites which act bidirectionally. Mol. Cell. Biol. 15, 5688-5699. https://doi.org/10.1128/MCB.15.10.5688
  20. Reddy, B.A., Etkin, L.D., and Freemont, P.S. 1992. A novel zinc finger coiled-coil domain in a family of nuclear proteins. Trends Biochem. Sci. 17, 344-345. https://doi.org/10.1016/0968-0004(92)90308-V
  21. Rosenfeld, M.G. 1991. POU-domain transcription factors: powerful developmental regulators. Genes Dev. 5, 897-907. https://doi.org/10.1101/gad.5.6.897
  22. Ruvkun, G. and Finney, M. 1991. Regulation of transcription and cell identity by POU domain proteins. Cell. 64, 475-478. https://doi.org/10.1016/0092-8674(91)90227-P
  23. Sambrook, J., Fritsch, E.F., and Maniatis, T. 1989. Molecular Cloning : A laboratory manual, 2nd ed. CSH.
  24. Smith, J., Anderson, J., Deans, S., and Davis, B. 1977. Asexual development in Aspergillus. pp. 23-58. In Smith, J.E. and Pateman, J.A. (eds.), Genetics and Physiology in Aspergillus. Academic Press, New York, N.Y., USA.
  25. Treisman, J., Harris, E., Wilson, D., and Desplan, C. 1992. The homeodomain: A new face for the helix-turn-helix. BioEssays 14, 145-150. https://doi.org/10.1002/bies.950140302
  26. Treisman, R. 1995. Inside the MADS box. Nature 376, 468-469. https://doi.org/10.1038/376468a0
  27. Yu, J.H., Wieser, J., and Adams,T.H. 1996. The Aspergillus FlbA RGS domain protein antagonizes G-protein signaling to block proliferation and allow development. EMBO J. 15, 5184-5190.
  28. Zonneveld, B.J.M. 1977. Biochemistry and ultrastructure of sexual development in Aspergillus nidulans. pp. 59-80. In Smith, J.E. and Pateman, J.A. (eds.), Genetics and Physiology in Aspergillus. Academic Press, New York, N.Y., USA.

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