Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1988 Apr;8(4):1504–1508. doi: 10.1128/mcb.8.4.1504

Molecular cloning and analysis of the regulation of cys-14+, a structural gene of the sulfur regulatory circuit of Neurospora crassa.

J S Ketter 1, G A Marzluf 1
PMCID: PMC363309  PMID: 2898097

Abstract

The cys-14+ gene encodes sulfate permease II, which is primarily expressed in mycelia. cys-14+ is one of a set of sulfur-related structural genes under the control of cys-3+ and scon+, the regulatory genes of the sulfur control circuit. We have cloned cys-14+ from a cosmid library of Neurospora crassa DNA. A restriction fragment length polymorphism analysis showed that this clone maps to the region of chromosome IV corresponding to the cys-14+ locus. Northern blot analyses were used to examine the regulated expression of the cys-14+ gene. In the wild type, a 3-kilobase cys-14+ transcript was highly expressed under sulfur-derepressing conditions but completely absent during sulfur repression. A cys-3 mutant, which cannot synthesize any of the sulfur-controlled enzymes, including sulfate permease II, did not possess any cys-14+ transcript under either condition. A cys-3 temperature-sensitive revertant completely lacked any cys-14+ mRNA at the conditional temperature but expressed the cys-14+ transcript upon derepression at the permissive temperature. Mutation of a second sulfur regulatory gene, scon(c), causes the expression of sulfur-related enzymes in a constitutive fashion; the scon(c) mutant showed a corresponding constitutive expression of cys-14+ mRNA, such that it was present even in cells subjected to sulfur repression conditions. These results show that the cys-14+ gene is regulated through the modulation of message content by the cys-3+ and scon(c) control genes in response to the sulfur levels of the cells.

Full text

PDF
1506

Images in this article

1505Image
on p.1505
1506Image
on p.1506

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. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burton E. G., Metzenberg R. L. Novel mutation causing derepression of several enzymes of sulfur metabolism in Neurospora crassa. J Bacteriol. 1972 Jan;109(1):140–151. doi: 10.1128/jb.109.1.140-151.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dietrich P. S., Metzenberg R. L. Metabolic suppressors of a regulatory mutant in Neurospora. Biochem Genet. 1973 Jan;8(1):73–84. doi: 10.1007/BF00485558. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Hanson M. A., Marzluf G. A. Regulation of a sulfur-controlled protease in Neurospora crassa. J Bacteriol. 1973 Nov;116(2):785–789. doi: 10.1128/jb.116.2.785-789.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Marzluf G. A. Control of the synthesis, activity, and turnover of enzymes of sulfur metabolism in Neurospora crassa. Arch Biochem Biophys. 1972 Jun;150(2):714–724. doi: 10.1016/0003-9861(72)90090-2. [DOI] [PubMed] [Google Scholar]
  9. Marzluf G. A. Genetic and biochemical studies of distinct sulfate permease species in different developmental stages of Neurospora crassa. Arch Biochem Biophys. 1970 May;138(1):254–263. doi: 10.1016/0003-9861(70)90306-1. [DOI] [PubMed] [Google Scholar]
  10. Marzluf G. A. Genetic and metabolic control of sulfate metabolism in Neurospora crassa: a specific permease for choline-O-sulfate. Biochem Genet. 1972 Dec;7(3):219–233. doi: 10.1007/BF00484820. [DOI] [PubMed] [Google Scholar]
  11. Marzluf G. A. Genetic and metabolic controls for sulfate metabolism in Neurospora crassa: isolation and study of chromate-resistant and sulfate transport-negative mutants. J Bacteriol. 1970 Jun;102(3):716–721. doi: 10.1128/jb.102.3.716-721.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Marzluf G. A., Metzenberg R. L. Positive control by the cys-3 locus in regulation of sulfur metabolism in Neurospora. J Mol Biol. 1968 Apr 28;33(2):423–437. doi: 10.1016/0022-2836(68)90199-x. [DOI] [PubMed] [Google Scholar]
  13. Metzenberg R. L., Ahlgren S. K. Mutants of Neurospora deficient in aryl sulfatase. Genetics. 1970 Mar-Apr;64(3):409–422. [PMC free article] [PubMed] [Google Scholar]
  14. Metzenberg R. L., Ahlgren S. K. Structural and regulatory control of aryl sulfatase in Neurospora: the use of interspecific differences in structural genes. Genetics. 1971 Jul;68(3):369–381. doi: 10.1093/genetics/68.3.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Metzenberg R. L., Parson J. W. Altered repression of some enzymes of sulfur utilization in a temperature-conditional lethal mutant of Neurospora. Proc Natl Acad Sci U S A. 1966 Mar;55(3):629–635. doi: 10.1073/pnas.55.3.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Orbach M. J., Porro E. B., Yanofsky C. Cloning and characterization of the gene for beta-tubulin from a benomyl-resistant mutant of Neurospora crassa and its use as a dominant selectable marker. Mol Cell Biol. 1986 Jul;6(7):2452–2461. doi: 10.1128/mcb.6.7.2452. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Pall M. L. Amino acid transport in Neurospora crassa. IV. Properties and regulation of a methionine transport system. Biochim Biophys Acta. 1971 Mar 9;233(1):201–214. doi: 10.1016/0005-2736(71)90372-5. [DOI] [PubMed] [Google Scholar]
  19. Reinert W. R., Marzluf G. A. Regulation of sulfate metabolism in Neurospora crassa: transport and accumulation of glucose 6-sulfate. Biochem Genet. 1974 Aug;12(2):97–108. doi: 10.1007/BF00487819. [DOI] [PubMed] [Google Scholar]
  20. Reinert W. R., Patel V. B., Giles N. H. Genetic regulation of the qa gene cluster of Neurospora crassa: induction of qa messenger ribonucleic acid and dependency on qa-1 function. Mol Cell Biol. 1981 Sep;1(9):829–835. doi: 10.1128/mcb.1.9.829. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  22. Schweizer M., Case M. E., Dykstra C. C., Giles N. H., Kushner S. R. Identification and characterization of recombinant plasmids carrying the complete qa gene cluster from Neurospora crassa including the qa-1+ regulatory gene. Proc Natl Acad Sci U S A. 1981 Aug;78(8):5086–5090. doi: 10.1073/pnas.78.8.5086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]

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

RESOURCES