Abstract
In Fusarium sporotrichioides, several genes required for biosynthesis of the trichothecene mycotoxin T-2 toxin are closely linked. Further characterization of this gene cluster has revealed a gene, Tri6, that specifies a 217-amino-acid protein with regions similar to Cys2His2 zinc finger proteins. Temporal expression of Tri6 is similar to that of trichothecene biosynthetic pathway genes. Analysis of Tri6 transcripts indicated that transcription is initiated in two regions and that within each region there may be at least four initiation sites. Disruption of Tri6 resulted in a mutant that did not produce trichothecenes but that did accumulate low levels of the trichothecene precursor trichodiene. The Tri6 mutant was unable to convert six trichothecene biosynthetic intermediates to T-2 toxin, and transcription of two biosynthetic genes, Tri4 and Tri5, was greatly reduced in the mutant relative to the wild type. In addition, the product of Tri6 functioned as a transcriptional activator in Saccharomyces cerevisiae when fused to the DNA binding region of GAL4. These results indicate that Tri6 encodes a protein involved in the transcriptional regulation of trichothecene biosynthetic genes in F. sporotrichioides.
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- 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]
- 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]
- Archambault J., Milne C. A., Schappert K. T., Baum B., Friesen J. D., Segall J. The deduced sequence of the transcription factor TFIIIA from Saccharomyces cerevisiae reveals extensive divergence from Xenopus TFIIIA. J Biol Chem. 1992 Feb 15;267(5):3282–3288. [PubMed] [Google Scholar]
- Asch D. K., Kinsey J. A. Relationship of vector insert size to homologous integration during transformation of Neurospora crassa with the cloned am (GDH) gene. Mol Gen Genet. 1990 Mar;221(1):37–43. doi: 10.1007/BF00280365. [DOI] [PubMed] [Google Scholar]
- Baldarelli R. M., Mahoney P. A., Salas F., Gustavson E., Boyer P. D., Chang M. F., Roark M., Lengyel J. A. Transcripts of the Drosophila blastoderm-specific locus, terminus, are concentrated posteriorly and encode a potential DNA-binding finger. Dev Biol. 1988 Jan;125(1):85–95. doi: 10.1016/0012-1606(88)90061-9. [DOI] [PubMed] [Google Scholar]
- Chang P. K., Cary J. W., Bhatnagar D., Cleveland T. E., Bennett J. W., Linz J. E., Woloshuk C. P., Payne G. A. Cloning of the Aspergillus parasiticus apa-2 gene associated with the regulation of aflatoxin biosynthesis. Appl Environ Microbiol. 1993 Oct;59(10):3273–3279. doi: 10.1128/aem.59.10.3273-3279.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chien C. T., Bartel P. L., Sternglanz R., Fields S. The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9578–9582. doi: 10.1073/pnas.88.21.9578. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coleman J. E. Zinc proteins: enzymes, storage proteins, transcription factors, and replication proteins. Annu Rev Biochem. 1992;61:897–946. doi: 10.1146/annurev.bi.61.070192.004341. [DOI] [PubMed] [Google Scholar]
- Covitz P. A., Herskowitz I., Mitchell A. P. The yeast RME1 gene encodes a putative zinc finger protein that is directly repressed by a1-alpha 2. Genes Dev. 1991 Nov;5(11):1982–1989. doi: 10.1101/gad.5.11.1982. [DOI] [PubMed] [Google Scholar]
- Cullen D., Smalley E. B., Caldwell R. W. New process for T-2 toxin production. Appl Environ Microbiol. 1982 Aug;44(2):371–375. doi: 10.1128/aem.44.2.371-375.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Edwards J. B., Delort J., Mallet J. Oligodeoxyribonucleotide ligation to single-stranded cDNAs: a new tool for cloning 5' ends of mRNAs and for constructing cDNA libraries by in vitro amplification. Nucleic Acids Res. 1991 Oct 11;19(19):5227–5232. doi: 10.1093/nar/19.19.5227. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Estruch F. The yeast putative transcriptional repressor RGM1 is a proline-rich zinc finger protein. Nucleic Acids Res. 1991 Sep 25;19(18):4873–4877. doi: 10.1093/nar/19.18.4873. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ginsberg A. M., King B. O., Roeder R. G. Xenopus 5S gene transcription factor, TFIIIA: characterization of a cDNA clone and measurement of RNA levels throughout development. Cell. 1984 Dec;39(3 Pt 2):479–489. doi: 10.1016/0092-8674(84)90455-0. [DOI] [PubMed] [Google Scholar]
- Hohn T. M., Beremand M. N. Regulation of Trichodiene Synthase in Fusarium sporotrichioides and Gibberella pulicaris (Fusarium sambucinum). Appl Environ Microbiol. 1989 Jun;55(6):1500–1503. doi: 10.1128/aem.55.6.1500-1503.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hohn T. M., Beremand P. D. Isolation and nucleotide sequence of a sesquiterpene cyclase gene from the trichothecene-producing fungus Fusarium sporotrichioides. Gene. 1989 Jun 30;79(1):131–138. doi: 10.1016/0378-1119(89)90098-x. [DOI] [PubMed] [Google Scholar]
- Hohn T. M., Desjardins A. E. Isolation and gene disruption of the Tox5 gene encoding trichodiene synthase in Gibberella pulicaris. Mol Plant Microbe Interact. 1992 May-Jun;5(3):249–256. doi: 10.1094/mpmi-5-249. [DOI] [PubMed] [Google Scholar]
- Hohn T. M., Desjardins A. E., McCormick S. P. Analysis of Tox5 gene expression in Gibberella pulicaris strains with different trichothecene production phenotypes. Appl Environ Microbiol. 1993 Aug;59(8):2359–2363. doi: 10.1128/aem.59.8.2359-2363.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hohn T. M., McCormick S. P., Desjardins A. E. Evidence for a gene cluster involving trichothecene-pathway biosynthetic genes in Fusarium sporotrichioides. Curr Genet. 1993 Oct;24(4):291–295. doi: 10.1007/BF00336778. [DOI] [PubMed] [Google Scholar]
- Hohn T. M., Vanmiddlesworth F. Purification and characterization of the sesquiterpene cyclase trichodiene synthetase from Fusarium sporotrichioides. Arch Biochem Biophys. 1986 Dec;251(2):756–761. doi: 10.1016/0003-9861(86)90386-3. [DOI] [PubMed] [Google Scholar]
- Keegan L., Gill G., Ptashne M. Separation of DNA binding from the transcription-activating function of a eukaryotic regulatory protein. Science. 1986 Feb 14;231(4739):699–704. doi: 10.1126/science.3080805. [DOI] [PubMed] [Google Scholar]
- MacCabe A. P., Riach M. B., Unkles S. E., Kinghorn J. R. The Aspergillus nidulans npeA locus consists of three contiguous genes required for penicillin biosynthesis. EMBO J. 1990 Jan;9(1):279–287. doi: 10.1002/j.1460-2075.1990.tb08106.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mathison L., Soliday C., Stepan T., Aldrich T., Rambosek J. Cloning, characterization, and use in strain improvement of the Cephalosporium acremonium gene cefG encoding acetyl transferase. Curr Genet. 1993 Jan;23(1):33–41. doi: 10.1007/BF00336747. [DOI] [PubMed] [Google Scholar]
- McCormick S. P., Taylor S. L., Plattner R. D., Beremand M. N. Bioconversion of possible T-2 toxin precursors by a mutant strain of Fusarium sporotrichioides NRRL 3299. Appl Environ Microbiol. 1990 Mar;56(3):702–706. doi: 10.1128/aem.56.3.702-706.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Payne G. A., Nystrom G. J., Bhatnagar D., Cleveland T. E., Woloshuk C. P. Cloning of the afl-2 gene involved in aflatoxin biosynthesis from Aspergillus flavus. Appl Environ Microbiol. 1993 Jan;59(1):156–162. doi: 10.1128/aem.59.1.156-162.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Párraga G., Horvath S., Hood L., Young E. T., Klevit R. E. Spectroscopic studies of wild-type and mutant "zinc finger" peptides: determinants of domain folding and structure. Proc Natl Acad Sci U S A. 1990 Jan;87(1):137–141. doi: 10.1073/pnas.87.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Salch Y. P., Beremand M. N. Gibberella pulicaris transformants: state of transforming DNA during asexual and sexual growth. Curr Genet. 1993;23(4):343–350. doi: 10.1007/BF00310897. [DOI] [PubMed] [Google Scholar]
- Salvato M. S., Shimomaye E. M. The completed sequence of lymphocytic choriomeningitis virus reveals a unique RNA structure and a gene for a zinc finger protein. Virology. 1989 Nov;173(1):1–10. doi: 10.1016/0042-6822(89)90216-x. [DOI] [PubMed] [Google Scholar]
- Salvato M., Shimomaye E., Southern P., Oldstone M. B. Virus-lymphocyte interactions. IV. Molecular characterization of LCMV Armstrong (CTL+) small genomic segment and that of its variant, Clone 13 (CTL-). Virology. 1988 Jun;164(2):517–522. doi: 10.1016/0042-6822(88)90566-1. [DOI] [PubMed] [Google Scholar]
- Schiestl R. H., Gietz R. D. High efficiency transformation of intact yeast cells using single stranded nucleic acids as a carrier. Curr Genet. 1989 Dec;16(5-6):339–346. doi: 10.1007/BF00340712. [DOI] [PubMed] [Google Scholar]
- Skory C. D., Chang P. K., Cary J., Linz J. E. Isolation and characterization of a gene from Aspergillus parasiticus associated with the conversion of versicolorin A to sterigmatocystin in aflatoxin biosynthesis. Appl Environ Microbiol. 1992 Nov;58(11):3527–3537. doi: 10.1128/aem.58.11.3527-3537.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Smith D. J., Burnham M. K., Edwards J., Earl A. J., Turner G. Cloning and heterologous expression of the penicillin biosynthetic gene cluster from penicillum chrysogenum. Biotechnology (N Y) 1990 Jan;8(1):39–41. doi: 10.1038/nbt0190-39. [DOI] [PubMed] [Google Scholar]
- Turgeon B. G., Garber R. C., Yoder O. C. Development of a fungal transformation system based on selection of sequences with promoter activity. Mol Cell Biol. 1987 Sep;7(9):3297–3305. doi: 10.1128/mcb.7.9.3297. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Valleley E. M., Müller U., Ferguson M. W., Sharpe P. T. Cloning and expression analysis of two ZFY-related zinc finger genes from Alligator mississippiensis, a species with temperature-dependent sex determination. Gene. 1992 Oct 1;119(2):221–228. doi: 10.1016/0378-1119(92)90275-t. [DOI] [PubMed] [Google Scholar]
- Vincent A., Colot H. V., Rosbash M. Sequence and structure of the serendipity locus of Drosophila melanogaster. A densely transcribed region including a blastoderm-specific gene. J Mol Biol. 1985 Nov 5;186(1):149–166. doi: 10.1016/0022-2836(85)90265-7. [DOI] [PubMed] [Google Scholar]