Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 2001 Mar;157(3):1003–1014. doi: 10.1093/genetics/157.3.1003

An STE12 homolog from the asexual, dimorphic fungus Penicillium marneffei complements the defect in sexual development of an Aspergillus nidulans steA mutant.

A R Borneman 1, M J Hynes 1, A Andrianopoulos 1
PMCID: PMC1461550  PMID: 11238390

Abstract

Penicillium marneffei is an opportunistic fungal pathogen of humans and the only dimorphic species identified in its genus. At 25 degrees P. marneffei exhibits true filamentous growth, while at 37 degrees P. marneffei undergoes a dimorphic transition to produce uninucleate yeast cells that divide by fission. Members of the STE12 family of regulators are involved in controlling mating and yeast-hyphal transitions in a number of fungi. We have cloned a homolog of the S. cerevisiae STE12 gene from P. marneffei, stlA, which is highly conserved. The stlA gene, along with the A. nidulans steA and Cryptococcus neoformans STE12alpha genes, form a distinct subclass of STE12 homologs that have a C2H2 zinc-finger motif in addition to the homeobox domain that defines STE12 genes. To examine the function of stlA in P. marneffei, we isolated a number of mutants in the P. marneffei-type strain and, in combination with selectable markers, developed a highly efficient DNA-mediated transformation procedure and gene deletion strategy. Deletion of the stlA gene had no detectable effect on vegetative growth, asexual development, or dimorphic switching in P. marneffei. Despite the lack of a detectable function, the P. marneffei stlA gene complemented the sexual defect of an A. nidulans steA mutant. In addition, substitution rate estimates indicate that there is a significant bias against nonsynonymous substitutions. These data suggest that P. marneffei may have a previously unidentified cryptic sexual cycle.

Full Text

The Full Text of this article is available as a PDF (568.8 KB).

Selected References

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

  1. Andrianopoulos A., Hynes M. J. Cloning and analysis of the positively acting regulatory gene amdR from Aspergillus nidulans. Mol Cell Biol. 1988 Aug;8(8):3532–3541. doi: 10.1128/mcb.8.8.3532-3541.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andrianopoulos A., Timberlake W. E. The Aspergillus nidulans abaA gene encodes a transcriptional activator that acts as a genetic switch to control development. Mol Cell Biol. 1994 Apr;14(4):2503–2515. doi: 10.1128/mcb.14.4.2503-2515.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Austin B., Hall R. M., Tyler B. M. Optimized vectors and selection for transformation of Neurospora crassa and Aspergillus nidulans to bleomycin and phleomycin resistance. Gene. 1990 Sep 1;93(1):157–162. doi: 10.1016/0378-1119(90)90152-h. [DOI] [PubMed] [Google Scholar]
  4. Borneman A. R., Hynes M. J., Andrianopoulos A. The abaA homologue of Penicillium marneffei participates in two developmental programmes: conidiation and dimorphic growth. Mol Microbiol. 2000 Dec;38(5):1034–1047. doi: 10.1046/j.1365-2958.2000.02202.x. [DOI] [PubMed] [Google Scholar]
  5. 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.7.9.3113-3118.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chan Y. F., Chow T. C. Ultrastructural observations on Penicillium marneffei in natural human infection. Ultrastruct Pathol. 1990 Sep-Oct;14(5):439–452. doi: 10.3109/01913129009007223. [DOI] [PubMed] [Google Scholar]
  7. Chang Y. C., Timberlake W. E. Identification of Aspergillus brlA response elements (BREs) by genetic selection in yeast. Genetics. 1993 Jan;133(1):29–38. doi: 10.1093/genetics/133.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chang Y. C., Wickes B. L., Miller G. F., Penoyer L. A., Kwon-Chung K. J. Cryptococcus neoformans STE12alpha regulates virulence but is not essential for mating. J Exp Med. 2000 Mar 6;191(5):871–882. doi: 10.1084/jem.191.5.871. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cobb M. H., Goldsmith E. J. How MAP kinases are regulated. J Biol Chem. 1995 Jun 23;270(25):14843–14846. doi: 10.1074/jbc.270.25.14843. [DOI] [PubMed] [Google Scholar]
  10. Cook J. G., Bardwell L., Thorner J. Inhibitory and activating functions for MAPK Kss1 in the S. cerevisiae filamentous-growth signalling pathway. Nature. 1997 Nov 6;390(6655):85–88. doi: 10.1038/36355. [DOI] [PubMed] [Google Scholar]
  11. Cooper C. R., Jr, Haycocks N. G. Penicillium marneffei: an insurgent species among the penicillia. J Eukaryot Microbiol. 2000 Jan-Feb;47(1):24–28. doi: 10.1111/j.1550-7408.2000.tb00006.x. [DOI] [PubMed] [Google Scholar]
  12. Cooper J. A. MAP kinase pathways. Straight and narrow or tortuous and intersecting? Curr Biol. 1994 Dec 1;4(12):1118–1121. doi: 10.1016/s0960-9822(00)00251-7. [DOI] [PubMed] [Google Scholar]
  13. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dutton J. R., Johns S., Miller B. L. StuAp is a sequence-specific transcription factor that regulates developmental complexity in Aspergillus nidulans. EMBO J. 1997 Sep 15;16(18):5710–5721. doi: 10.1093/emboj/16.18.5710. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ehinger A., Denison S. H., May G. S. Sequence, organization and expression of the core histone genes of Aspergillus nidulans. Mol Gen Genet. 1990 Jul;222(2-3):416–424. doi: 10.1007/BF00633848. [DOI] [PubMed] [Google Scholar]
  16. Fields S., Herskowitz I. Regulation by the yeast mating-type locus of STE12, a gene required for cell-type-specific expression. Mol Cell Biol. 1987 Oct;7(10):3818–3821. doi: 10.1128/mcb.7.10.3818-3821.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Fincham J. R. Transformation in fungi. Microbiol Rev. 1989 Mar;53(1):148–170. doi: 10.1128/mr.53.1.148-170.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Garrison R. G., Boyd K. S. Dimorphism of Penicillium marneffei as observed by electron microscopy. Can J Microbiol. 1973 Oct;19(10):1305–1309. doi: 10.1139/m73-209. [DOI] [PubMed] [Google Scholar]
  19. Geiser D. M., Timberlake W. E., Arnold M. L. Loss of meiosis in Aspergillus. Mol Biol Evol. 1996 Jul;13(6):809–817. doi: 10.1093/oxfordjournals.molbev.a025641. [DOI] [PubMed] [Google Scholar]
  20. Gustin M. C., Albertyn J., Alexander M., Davenport K. MAP kinase pathways in the yeast Saccharomyces cerevisiae. Microbiol Mol Biol Rev. 1998 Dec;62(4):1264–1300. doi: 10.1128/mmbr.62.4.1264-1300.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hull C. M., Raisner R. M., Johnson A. D. Evidence for mating of the "asexual" yeast Candida albicans in a mammalian host. Science. 2000 Jul 14;289(5477):307–310. doi: 10.1126/science.289.5477.307. [DOI] [PubMed] [Google Scholar]
  22. Kronstad J. W., Holly J. A., MacKay V. L. A yeast operator overlaps an upstream activation site. Cell. 1987 Jul 31;50(3):369–377. doi: 10.1016/0092-8674(87)90491-0. [DOI] [PubMed] [Google Scholar]
  23. Kwon-Chung K. J., Bennett J. E. Distribution of alpha and alpha mating types of Cryptococcus neoformans among natural and clinical isolates. Am J Epidemiol. 1978 Oct;108(4):337–340. doi: 10.1093/oxfordjournals.aje.a112628. [DOI] [PubMed] [Google Scholar]
  24. Kwon-Chung K. J., Edman J. C., Wickes B. L. Genetic association of mating types and virulence in Cryptococcus neoformans. Infect Immun. 1992 Feb;60(2):602–605. doi: 10.1128/iai.60.2.602-605.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Leberer E., Harcus D., Broadbent I. D., Clark K. L., Dignard D., Ziegelbauer K., Schmidt A., Gow N. A., Brown A. J., Thomas D. Y. Signal transduction through homologs of the Ste20p and Ste7p protein kinases can trigger hyphal formation in the pathogenic fungus Candida albicans. Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):13217–13222. doi: 10.1073/pnas.93.23.13217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lo H. J., Köhler J. R., DiDomenico B., Loebenberg D., Cacciapuoti A., Fink G. R. Nonfilamentous C. albicans mutants are avirulent. Cell. 1997 Sep 5;90(5):939–949. doi: 10.1016/s0092-8674(00)80358-x. [DOI] [PubMed] [Google Scholar]
  27. Madhani H. D., Fink G. R. Combinatorial control required for the specificity of yeast MAPK signaling. Science. 1997 Feb 28;275(5304):1314–1317. doi: 10.1126/science.275.5304.1314. [DOI] [PubMed] [Google Scholar]
  28. Madhani H. D., Fink G. R. The riddle of MAP kinase signaling specificity. Trends Genet. 1998 Apr;14(4):151–155. doi: 10.1016/s0168-9525(98)01425-5. [DOI] [PubMed] [Google Scholar]
  29. Madhani H. D., Styles C. A., Fink G. R. MAP kinases with distinct inhibitory functions impart signaling specificity during yeast differentiation. Cell. 1997 Nov 28;91(5):673–684. doi: 10.1016/s0092-8674(00)80454-7. [DOI] [PubMed] [Google Scholar]
  30. Miller K. Y., Wu J., Miller B. L. StuA is required for cell pattern formation in Aspergillus. Genes Dev. 1992 Sep;6(9):1770–1782. doi: 10.1101/gad.6.9.1770. [DOI] [PubMed] [Google Scholar]
  31. Oakley B. R., Rinehart J. E., Mitchell B. L., Oakley C. E., Carmona C., Gray G. L., May G. S. Cloning, mapping and molecular analysis of the pyrG (orotidine-5'-phosphate decarboxylase) gene of Aspergillus nidulans. Gene. 1987;61(3):385–399. doi: 10.1016/0378-1119(87)90201-0. [DOI] [PubMed] [Google Scholar]
  32. Pi H., Chien C. T., Fields S. Transcriptional activation upon pheromone stimulation mediated by a small domain of Saccharomyces cerevisiae Ste12p. Mol Cell Biol. 1997 Nov;17(11):6410–6418. doi: 10.1128/mcb.17.11.6410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Raymond M., Dignard D., Alarco A. M., Mainville N., Magee B. B., Thomas D. Y. A Ste6p/P-glycoprotein homologue from the asexual yeast Candida albicans transports the a-factor mating pheromone in Saccharomyces cerevisiae. Mol Microbiol. 1998 Feb;27(3):587–598. doi: 10.1046/j.1365-2958.1998.00704.x. [DOI] [PubMed] [Google Scholar]
  34. Sadhu C., Hoekstra D., McEachern M. J., Reed S. I., Hicks J. B. A G-protein alpha subunit from asexual Candida albicans functions in the mating signal transduction pathway of Saccharomyces cerevisiae and is regulated by the a1-alpha 2 repressor. Mol Cell Biol. 1992 May;12(5):1977–1985. doi: 10.1128/mcb.12.5.1977-1985.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Song D., Dolan J. W., Yuan Y. L., Fields S. Pheromone-dependent phosphorylation of the yeast STE12 protein correlates with transcriptional activation. Genes Dev. 1991 May;5(5):741–750. doi: 10.1101/gad.5.5.741. [DOI] [PubMed] [Google Scholar]
  36. Suelmann R., Sievers N., Fischer R. Nuclear traffic in fungal hyphae: in vivo study of nuclear migration and positioning in Aspergillus nidulans. Mol Microbiol. 1997 Aug;25(4):757–769. doi: 10.1046/j.1365-2958.1997.5131873.x. [DOI] [PubMed] [Google Scholar]
  37. 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]
  38. Vallim M. A., Miller K. Y., Miller B. L. Aspergillus SteA (sterile12-like) is a homeodomain-C2/H2-Zn+2 finger transcription factor required for sexual reproduction. Mol Microbiol. 2000 Apr;36(2):290–301. doi: 10.1046/j.1365-2958.2000.01874.x. [DOI] [PubMed] [Google Scholar]
  39. Wickes B. L., Edman U., Edman J. C. The Cryptococcus neoformans STE12alpha gene: a putative Saccharomyces cerevisiae STE12 homologue that is mating type specific. Mol Microbiol. 1997 Dec;26(5):951–960. doi: 10.1046/j.1365-2958.1997.6322001.x. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. Young L. Y., Lorenz M. C., Heitman J. A STE12 homolog is required for mating but dispensable for filamentation in candida lusitaniae. Genetics. 2000 May;155(1):17–29. doi: 10.1093/genetics/155.1.17. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Yuan Y. L., Fields S. Properties of the DNA-binding domain of the Saccharomyces cerevisiae STE12 protein. Mol Cell Biol. 1991 Dec;11(12):5910–5918. doi: 10.1128/mcb.11.12.5910-5918.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Yue C., Cavallo L. M., Alspaugh J. A., Wang P., Cox G. M., Perfect J. R., Heitman J. The STE12alpha homolog is required for haploid filamentation but largely dispensable for mating and virulence in Cryptococcus neoformans. Genetics. 1999 Dec;153(4):1601–1615. doi: 10.1093/genetics/153.4.1601. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES