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. 1990 Nov;10(11):5839–5848. doi: 10.1128/mcb.10.11.5839

Molecular analysis of nuc-1+, a gene controlling phosphorus acquisition in Neurospora crassa.

S Kang 1, R L Metzenberg 1
PMCID: PMC361368  PMID: 2146493

Abstract

In response to phosphorus starvation, Neurospora crassa makes several enzymes that are undetectable or barely detectable in phosphate-sufficient cultures. The nuc-1+ gene, whose product regulates the synthesis of these enzymes, was cloned and sequenced. The nuc-1+ gene encodes a protein of 824 amino acids with a predicted molecular weight of 87,429. The amino acid sequence shows homology with two yeast proteins whose functions are analogous to that of the NUC-1 protein. Two nuc-1+ transcripts of 3.2 and 3.0 kilobases were detected; they were present in similar amounts during growth at low or high phosphate concentrations. The nuc-2+ gene encodes a product normally required for NUC-1 function, and yet a nuc-2 mutation can be complemented by overexpression of the nuc-1+ gene. This implies physical interactions between NUC-1 protein and the negative regulatory factor(s) PREG and/or PGOV. Analysis of nuc-2 and nuc-1; nuc-2 strains transformed by the nuc-1+ gene suggests that phosphate directly affects the level or activity of the negative regulatory factor(s) controlling phosphorus acquisition.

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Selected References

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

  1. Akins R. A., Lambowitz A. M. General method for cloning Neurospora crassa nuclear genes by complementation of mutants. Mol Cell Biol. 1985 Sep;5(9):2272–2278. doi: 10.1128/mcb.5.9.2272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arndt K. T., Styles C., Fink G. R. Multiple global regulators control HIS4 transcription in yeast. Science. 1987 Aug 21;237(4817):874–880. doi: 10.1126/science.3303332. [DOI] [PubMed] [Google Scholar]
  3. Barthelmess I. B. Mutants affecting amino acid cross-pathway control in Neurospora crassa. Genet Res. 1982 Apr;39(2):169–185. doi: 10.1017/s0016672300020863. [DOI] [PubMed] [Google Scholar]
  4. Courey A. J., Tjian R. Analysis of Sp1 in vivo reveals multiple transcriptional domains, including a novel glutamine-rich activation motif. Cell. 1988 Dec 2;55(5):887–898. doi: 10.1016/0092-8674(88)90144-4. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  7. Fu Y. H., Marzluf G. A. Characterization of nit-2, the major nitrogen regulatory gene of Neurospora crassa. Mol Cell Biol. 1987 May;7(5):1691–1696. doi: 10.1128/mcb.7.5.1691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fu Y. H., Paietta J. V., Mannix D. G., Marzluf G. A. cys-3, the positive-acting sulfur regulatory gene of Neurospora crassa, encodes a protein with a putative leucine zipper DNA-binding element. Mol Cell Biol. 1989 Mar;9(3):1120–1127. doi: 10.1128/mcb.9.3.1120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Geever R. F., Huiet L., Baum J. A., Tyler B. M., Patel V. B., Rutledge B. J., Case M. E., Giles N. H. DNA sequence, organization and regulation of the qa gene cluster of Neurospora crassa. J Mol Biol. 1989 May 5;207(1):15–34. doi: 10.1016/0022-2836(89)90438-5. [DOI] [PubMed] [Google Scholar]
  10. Hasunuma K. Repressible extracellular nucleases in Neurospora crassa. Biochim Biophys Acta. 1973 Sep 7;319(3):288–293. doi: 10.1016/0005-2787(73)90168-8. [DOI] [PubMed] [Google Scholar]
  11. Johnston M. A model fungal gene regulatory mechanism: the GAL genes of Saccharomyces cerevisiae. Microbiol Rev. 1987 Dec;51(4):458–476. doi: 10.1128/mr.51.4.458-476.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kammerer B., Guyonvarch A., Hubert J. C. Yeast regulatory gene PPR1. I. Nucleotide sequence, restriction map and codon usage. J Mol Biol. 1984 Dec 5;180(2):239–250. doi: 10.1016/s0022-2836(84)80002-9. [DOI] [PubMed] [Google Scholar]
  13. Kinsey J. A., Rambosek J. A. Transformation of Neurospora crassa with the cloned am (glutamate dehydrogenase) gene. Mol Cell Biol. 1984 Jan;4(1):117–122. doi: 10.1128/mcb.4.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Legrain M., De Wilde M., Hilger F. Isolation, physical characterization and expression analysis of the Saccharomyces cerevisiae positive regulatory gene PHO4. Nucleic Acids Res. 1986 Apr 11;14(7):3059–3073. doi: 10.1093/nar/14.7.3059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lehman J. F., Gleason M. K., Ahlgren S. K., Metzenberg R. L. Regulation of phosphate metabolism in Neurospora crassa. Characterization of regulatory mutants. Genetics. 1973 Sep;75(1):61–73. doi: 10.1093/genetics/75.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lowendorf H. S., Slayman C. W. Genetic regulation of phosphate transport system II in Neurospora. Biochim Biophys Acta. 1975 Nov 17;413(1):95–103. doi: 10.1016/0005-2736(75)90061-9. [DOI] [PubMed] [Google Scholar]
  17. Mann B. J., Akins R. A., Lambowitz A. M., Metzenberg R. L. The structural gene for a phosphorus-repressible phosphate permease in Neurospora crassa can complement a mutation in positive regulatory gene nuc-1. Mol Cell Biol. 1988 Mar;8(3):1376–1379. doi: 10.1128/mcb.8.3.1376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mann B. J., Bowman B. J., Grotelueschen J., Metzenberg R. L. Nucleotide sequence of pho-4+, encoding a phosphate-repressible phosphate permease of Neurospora crassa. Gene. 1989 Nov 30;83(2):281–289. doi: 10.1016/0378-1119(89)90114-5. [DOI] [PubMed] [Google Scholar]
  19. Mermod N., O'Neill E. A., Kelly T. J., Tjian R. The proline-rich transcriptional activator of CTF/NF-I is distinct from the replication and DNA binding domain. Cell. 1989 Aug 25;58(4):741–753. doi: 10.1016/0092-8674(89)90108-6. [DOI] [PubMed] [Google Scholar]
  20. Metzenberg R. L., Chia W. Genetic control of phosphorus assimilation in Neurospora crassa: dose-dependent dominance and recessiveness in constitutive mutants. Genetics. 1979 Nov;93(3):625–643. doi: 10.1093/genetics/93.3.625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mitchell P. J., Tjian R. Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. Science. 1989 Jul 28;245(4916):371–378. doi: 10.1126/science.2667136. [DOI] [PubMed] [Google Scholar]
  22. Murre C., McCaw P. S., Baltimore D. A new DNA binding and dimerization motif in immunoglobulin enhancer binding, daughterless, MyoD, and myc proteins. Cell. 1989 Mar 10;56(5):777–783. doi: 10.1016/0092-8674(89)90682-x. [DOI] [PubMed] [Google Scholar]
  23. Nyc J. F., Kadner R. J., Crocken B. J. A repressible alkaline phosphatase in Neurospora crassa. J Biol Chem. 1966 Apr 10;241(7):1468–1472. [PubMed] [Google Scholar]
  24. Ogawa N., Oshima Y. Functional domains of a positive regulatory protein, PHO4, for transcriptional control of the phosphatase regulon in Saccharomyces cerevisiae. Mol Cell Biol. 1990 May;10(5):2224–2236. doi: 10.1128/mcb.10.5.2224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Paietta J. V., Akins R. A., Lambowitz A. M., Marzluf G. A. Molecular cloning and characterization of the cys-3 regulatory gene of Neurospora crassa. Mol Cell Biol. 1987 Jul;7(7):2506–2511. doi: 10.1128/mcb.7.7.2506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Patel V. B., Giles N. H. Autogenous regulation of the positive regulatory qa-1F gene in Neurospora crassa. Mol Cell Biol. 1985 Dec;5(12):3593–3599. doi: 10.1128/mcb.5.12.3593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Perkins D. D., Barry E. G. The cytogenetics of Neurospora. Adv Genet. 1977;19:133–285. doi: 10.1016/s0065-2660(08)60246-1. [DOI] [PubMed] [Google Scholar]
  28. Sengstag C., Hinnen A. The sequence of the Saccharomyces cerevisiae gene PHO2 codes for a regulatory protein with unusual aminoacid composition. Nucleic Acids Res. 1987 Jan 12;15(1):233–246. doi: 10.1093/nar/15.1.233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Toh-E A., Ishikawa T. Genetic control of the synthesis of repressible phosphatases in Neurospora crassa. Genetics. 1971 Nov;69(3):339–351. doi: 10.1093/genetics/69.3.339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Vogel K., Hörz W., Hinnen A. The two positively acting regulatory proteins PHO2 and PHO4 physically interact with PHO5 upstream activation regions. Mol Cell Biol. 1989 May;9(5):2050–2057. doi: 10.1128/mcb.9.5.2050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vollmer S. J., Yanofsky C. Efficient cloning of genes of Neurospora crassa. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4869–4873. doi: 10.1073/pnas.83.13.4869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yoshida K., Kuromitsu Z., Ogawa N., Oshima Y. Mode of expression of the positive regulatory genes PHO2 and PHO4 of the phosphatase regulon in Saccharomyces cerevisiae. Mol Gen Genet. 1989 May;217(1):31–39. doi: 10.1007/BF00330939. [DOI] [PubMed] [Google Scholar]
  33. Yoshida K., Ogawa N., Oshima Y. Function of the PHO regulatory genes for repressible acid phosphatase synthesis in Saccharomyces cerevisiae. Mol Gen Genet. 1989 May;217(1):40–46. doi: 10.1007/BF00330940. [DOI] [PubMed] [Google Scholar]

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