Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1993 Nov;13(11):7091–7100. doi: 10.1128/mcb.13.11.7091

urbs1, a gene regulating siderophore biosynthesis in Ustilago maydis, encodes a protein similar to the erythroid transcription factor GATA-1.

C Voisard 1, J Wang 1, J L McEvoy 1, P Xu 1, S A Leong 1
PMCID: PMC364770  PMID: 8413298

Abstract

Ustilago maydis secretes ferrichrome-type siderophores, ferric-ion-binding compounds, in response to iron starvation. TA2701, a non-enterobactin-producing, non-ferrichrome-utilizing mutant of Salmonella typhimurium LT-2, was employed as a biological indicator in a novel screening method to isolate three N-methyl-N'-nitro-N-nitrosoguanidine-induced U. maydis mutants defective in the regulation of ferrichrome-type siderophore biosynthesis. These mutants displayed a constitutive phenotype; they produced siderophores in the presence of iron concentrations that would typically repress siderophore synthesis in wild-type strains. A 4.8-kb fragment of U. maydis genomic DNA capable of restoring normal regulation of siderophore biosynthesis in the constitutive mutants was identified. This segment of DNA contains an intronless open reading frame that specifies a protein of 950 amino acids containing two finger motifs similar to those found in the erythroid transcription factor GATA-1. Disruption of this open reading frame in a wild-type strain gave rise to cells that produced siderophores constitutively. Genetic studies indicated that the disruption mutation was allelic to the chemically induced mutations, confirming that the structural gene for a regulator rather than a suppressor gene had been cloned. Northern (RNA) analysis of the gene revealed a 4.2-kb transcript that is expressed constitutively at low levels in wild-type cells. The data support the hypothesis that this gene, which we designate urbs1 (Ustilago regulator of biosynthesis of siderophores), acts directly or indirectly to repress biosynthesis of siderophores in U. maydis.

Full text

PDF
7100

Images in this article

7098Image
on p.7098

Selected References

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

  1. Arnold F. H., Haymore B. L. Engineered metal-binding proteins: purification to protein folding. Science. 1991 Jun 28;252(5014):1796–1797. doi: 10.1126/science.1648261. [DOI] [PubMed] [Google Scholar]
  2. Atkin C. L., Neilands J. B., Phaff H. J. Rhodotorulic acid from species of Leucosporidium, Rhodosporidium, Rhodotorula, Sporidiobolus, and Sporobolomyces, and a new alanine-containing ferrichrome from Cryptococcus melibiosum. J Bacteriol. 1970 Sep;103(3):722–733. doi: 10.1128/jb.103.3.722-733.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bagg A., Neilands J. B. Molecular mechanism of regulation of siderophore-mediated iron assimilation. Microbiol Rev. 1987 Dec;51(4):509–518. doi: 10.1128/mr.51.4.509-518.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Budde A. D., Leong S. A. Characterization of siderophores from Ustilago maydis. Mycopathologia. 1989 Nov;108(2):125–133. doi: 10.1007/BF00436063. [DOI] [PubMed] [Google Scholar]
  5. Bysani N., Daugherty J. R., Cooper T. G. Saturation mutagenesis of the UASNTR (GATAA) responsible for nitrogen catabolite repression-sensitive transcriptional activation of the allantoin pathway genes in Saccharomyces cerevisiae. J Bacteriol. 1991 Aug;173(16):4977–4982. doi: 10.1128/jb.173.16.4977-4982.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Coy M., Neilands J. B. Structural dynamics and functional domains of the fur protein. Biochemistry. 1991 Aug 20;30(33):8201–8210. doi: 10.1021/bi00247a016. [DOI] [PubMed] [Google Scholar]
  7. Cunningham T. S., Cooper T. G. Expression of the DAL80 gene, whose product is homologous to the GATA factors and is a negative regulator of multiple nitrogen catabolic genes in Saccharomyces cerevisiae, is sensitive to nitrogen catabolite repression. Mol Cell Biol. 1991 Dec;11(12):6205–6215. doi: 10.1128/mcb.11.12.6205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Daum H. A., 3rd, White H. W., Seidell C. M., Johnson P. A. Cloning, restriction digestion and DNA labeling of large DNA fragments (greater than or equal to 1 kb) in the presence of remelted SeaPlaque GTG agarose gels. Biotechniques. 1991 Dec;11(6):784-6, 788, 790-1. [PubMed] [Google Scholar]
  9. 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]
  10. Durkin M. E., Chakravarti S., Bartos B. B., Liu S. H., Friedman R. L., Chung A. E. Amino acid sequence and domain structure of entactin. Homology with epidermal growth factor precursor and low density lipoprotein receptor. J Cell Biol. 1988 Dec;107(6 Pt 2):2749–2756. doi: 10.1083/jcb.107.6.2749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Elder R. T., Loh E. Y., Davis R. W. RNA from the yeast transposable element Ty1 has both ends in the direct repeats, a structure similar to retrovirus RNA. Proc Natl Acad Sci U S A. 1983 May;80(9):2432–2436. doi: 10.1073/pnas.80.9.2432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Frohman M. A., Dush M. K., Martin G. R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8998–9002. doi: 10.1073/pnas.85.23.8998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fu Y. H., Marzluf G. A. nit-2, the major positive-acting nitrogen regulatory gene of Neurospora crassa, encodes a sequence-specific DNA-binding protein. Proc Natl Acad Sci U S A. 1990 Jul;87(14):5331–5335. doi: 10.1073/pnas.87.14.5331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fuchs R., Stoehr P., Rice P., Omond R., Cameron G. New services of the EMBL Data Library. Nucleic Acids Res. 1990 Aug 11;18(15):4319–4323. doi: 10.1093/nar/18.15.4319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Halliwell B., Gutteridge J. M. Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J. 1984 Apr 1;219(1):1–14. doi: 10.1042/bj2190001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hannon R., Evans T., Felsenfeld G., Gould H. Structure and promoter activity of the gene for the erythroid transcription factor GATA-1. Proc Natl Acad Sci U S A. 1991 Apr 15;88(8):3004–3008. doi: 10.1073/pnas.88.8.3004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Holden D. W., Kronstad J. W., Leong S. A. Mutation in a heat-regulated hsp70 gene of Ustilago maydis. EMBO J. 1989 Jul;8(7):1927–1934. doi: 10.1002/j.1460-2075.1989.tb03596.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kronstad J. W., Leong S. A. The b mating-type locus of Ustilago maydis contains variable and constant regions. Genes Dev. 1990 Aug;4(8):1384–1395. doi: 10.1101/gad.4.8.1384. [DOI] [PubMed] [Google Scholar]
  19. Kudla B., Caddick M. X., Langdon T., Martinez-Rossi N. M., Bennett C. F., Sibley S., Davies R. W., Arst H. N., Jr The regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans. Mutations affecting specificity of gene activation alter a loop residue of a putative zinc finger. EMBO J. 1990 May;9(5):1355–1364. doi: 10.1002/j.1460-2075.1990.tb08250.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Köhrer K., Domdey H. Preparation of high molecular weight RNA. Methods Enzymol. 1991;194:398–405. doi: 10.1016/0076-6879(91)94030-g. [DOI] [PubMed] [Google Scholar]
  21. Lee S. Y., Rasheed S. A simple procedure for maximum yield of high-quality plasmid DNA. Biotechniques. 1990 Dec;9(6):676–679. [PubMed] [Google Scholar]
  22. Llinás M., Klein M. P., Neilands J. B. Solution conformation of ferrichrome, a microbial iron transport cyclohexapeptide, as deduced by high resolution proton magnetic resonance. J Mol Biol. 1970 Sep 28;52(3):399–414. doi: 10.1016/0022-2836(70)90409-2. [DOI] [PubMed] [Google Scholar]
  23. Luckey M., Pollack J. R., Wayne R., Ames B. N., Neilands J. B. Iron uptake in Salmonella typhimurium: utilization of exogenous siderochromes as iron carriers. J Bacteriol. 1972 Sep;111(3):731–738. doi: 10.1128/jb.111.3.731-738.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mei B., Budde A. D., Leong S. A. sid1, a gene initiating siderophore biosynthesis in Ustilago maydis: molecular characterization, regulation by iron, and role in phytopathogenicity. Proc Natl Acad Sci U S A. 1993 Feb 1;90(3):903–907. doi: 10.1073/pnas.90.3.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Minehart P. L., Magasanik B. Sequence and expression of GLN3, a positive nitrogen regulatory gene of Saccharomyces cerevisiae encoding a protein with a putative zinc finger DNA-binding domain. Mol Cell Biol. 1991 Dec;11(12):6216–6228. doi: 10.1128/mcb.11.12.6216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mlodzik M., Gehring W. J. Expression of the caudal gene in the germ line of Drosophila: formation of an RNA and protein gradient during early embryogenesis. Cell. 1987 Feb 13;48(3):465–478. doi: 10.1016/0092-8674(87)90197-8. [DOI] [PubMed] [Google Scholar]
  27. O'Hara P. J., Horowitz H., Eichinger G., Young E. T. The yeast ADR6 gene encodes homopolymeric amino acid sequences and a potential metal-binding domain. Nucleic Acids Res. 1988 Nov 11;16(21):10153–10169. doi: 10.1093/nar/16.21.10153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Omichinski J. G., Trainor C., Evans T., Gronenborn A. M., Clore G. M., Felsenfeld G. A small single-"finger" peptide from the erythroid transcription factor GATA-1 binds specifically to DNA as a zinc or iron complex. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1676–1680. doi: 10.1073/pnas.90.5.1676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Puhalla J. E. Compatibility reactions on solid medium and interstrain inhibition in Ustilago maydis. Genetics. 1968 Nov;60(3):461–474. doi: 10.1093/genetics/60.3.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sawadogo M., Van Dyke M. W. A rapid method for the purification of deprotected oligodeoxynucleotides. Nucleic Acids Res. 1991 Feb 11;19(3):674–674. doi: 10.1093/nar/19.3.674. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Schäffer S., Hantke K., Braun V. Nucleotide sequence of the iron regulatory gene fur. Mol Gen Genet. 1985;200(1):110–113. doi: 10.1007/BF00383321. [DOI] [PubMed] [Google Scholar]
  33. Smith T. L., Leong S. A. Isolation and characterization of a Ustilago maydis glyceraldehyde-3-phosphate dehydrogenase-encoding gene. Gene. 1990 Sep 1;93(1):111–117. doi: 10.1016/0378-1119(90)90143-f. [DOI] [PubMed] [Google Scholar]
  34. Suzuki M. SPXX, a frequent sequence motif in gene regulatory proteins. J Mol Biol. 1989 May 5;207(1):61–84. doi: 10.1016/0022-2836(89)90441-5. [DOI] [PubMed] [Google Scholar]
  35. Toneguzzo F., Glynn S., Levi E., Mjolsness S., Hayday A. Use of a chemically modified T7 DNA polymerase for manual and automated sequencing of supercoiled DNA. Biotechniques. 1988 May;6(5):460–469. [PubMed] [Google Scholar]
  36. Trainor C. D., Evans T., Felsenfeld G., Boguski M. S. Structure and evolution of a human erythroid transcription factor. Nature. 1990 Jan 4;343(6253):92–96. doi: 10.1038/343092a0. [DOI] [PubMed] [Google Scholar]
  37. Tsai S. F., Martin D. I., Zon L. I., D'Andrea A. D., Wong G. G., Orkin S. H. Cloning of cDNA for the major DNA-binding protein of the erythroid lineage through expression in mammalian cells. Nature. 1989 Jun 8;339(6224):446–451. doi: 10.1038/339446a0. [DOI] [PubMed] [Google Scholar]
  38. Tsukuda T., Carleton S., Fotheringham S., Holloman W. K. Isolation and characterization of an autonomously replicating sequence from Ustilago maydis. Mol Cell Biol. 1988 Sep;8(9):3703–3709. doi: 10.1128/mcb.8.9.3703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wang J., Budde A. D., Leong S. A. Analysis of ferrichrome biosynthesis in the phytopathogenic fungus Ustilago maydis: cloning of an ornithine-N5-oxygenase gene. J Bacteriol. 1989 May;171(5):2811–2818. doi: 10.1128/jb.171.5.2811-2818.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wang J., Holden D. W., Leong S. A. Gene transfer system for the phytopathogenic fungus Ustilago maydis. Proc Natl Acad Sci U S A. 1988 Feb;85(3):865–869. doi: 10.1073/pnas.85.3.865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Zheng L., Andrews P. C., Hermodson M. A., Dixon J. E., Zalkin H. Cloning and structural characterization of porcine heart aconitase. J Biol Chem. 1990 Feb 15;265(5):2814–2821. [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES