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. 1990 Mar;124(3):599–606. doi: 10.1093/genetics/124.3.599

Determination of the Inactivating Alterations in Two Mutant Alleles of the Neurospora Crassa Cross-Pathway Control Gene Cpc-1

J L Paluh 1, M Plamann 1, D Kruger 1, I B Barthelmess 1, C Yanofsky 1, D D Perkins 1
PMCID: PMC1203953  PMID: 2138111

Abstract

cpc-1 is the locus specifying what is believed to be the major trans-activating transcription factor that regulates expression of amino acid biosynthetic genes subject to cross-pathway control in Neurospora crassa. Mutants altered at this locus are incapable of the global increase in gene expression normally seen in response to amino acid starvation. Using polymerase chain reaction methodology we have cloned and sequenced the inactive mutant allele, cpc-1 (CD15). The cpc-1 (CD15) mutation was found to be a single base pair deletion in codon 93 of the cpc-1 structural gene. A second, presumed lethal, allele, cpc-1 (j-5), also was investigated. Northern analyses with strains carrying the cpc-1 (j-5) allele revealed that no cpc-1 mRNA is produced. Southern and genetic analyses established that the cpc-1 (j-5) mutation involved a chromosomal rearrangement in which a break occurred within the cpc-1 locus, normally resident on linkage group VI; a small fragment from the left arm of linkage group VI, containing the cpc-1 promoter region and ylo-1, was translocated to the right arm of linkage group I. Other studies indicate that the cpc-1 locus itself is not essential for viability. Lethality previously attributed to the cpc-1 (j-5) mutation is due instead to the production of progeny that are deficient for essential genes in an adjoining segment of linkage group VI. Molecular characterization of cpc-1 (j-5) X ylo-1 pan-2 duplication progeny indicated that cpc-1 is normally transcribed towards the linkage group VI centromere.

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Selected References

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

  1. 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]
  2. Carsiotis M., Jones R. F. Cross-pathway regulation: tryptophan-mediated control of histidine and arginine biosynthetic enzymes in Neurospora crassa. J Bacteriol. 1974 Sep;119(3):889–892. doi: 10.1128/jb.119.3.889-892.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carsiotis M., Jones R. F., Wesseling A. C. Cross-pathway regulation: histidine-mediated control of histidine, tryptophan, and arginine biosynthetic enzymes in Neurospora crassa. J Bacteriol. 1974 Sep;119(3):893–898. doi: 10.1128/jb.119.3.893-898.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Flint H. J., Wilkening J. Cloning of the arg-12 gene of Neurospora crassa and regulation of its transcript via cross-pathway amino acid control. Mol Gen Genet. 1986 Apr;203(1):110–116. doi: 10.1007/BF00330391. [DOI] [PubMed] [Google Scholar]
  6. Goc A., Wegleński P. Regulatory region of the Aspergillus nidulans argB gene. Curr Genet. 1988 Nov;14(5):425–429. doi: 10.1007/BF00521264. [DOI] [PubMed] [Google Scholar]
  7. Hinnebusch A. G. Mechanisms of gene regulation in the general control of amino acid biosynthesis in Saccharomyces cerevisiae. Microbiol Rev. 1988 Jun;52(2):248–273. doi: 10.1128/mr.52.2.248-273.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hope I. A., Struhl K. GCN4 protein, synthesized in vitro, binds HIS3 regulatory sequences: implications for general control of amino acid biosynthetic genes in yeast. Cell. 1985 Nov;43(1):177–188. doi: 10.1016/0092-8674(85)90022-4. [DOI] [PubMed] [Google Scholar]
  9. Hope I. A., Struhl K. GCN4, a eukaryotic transcriptional activator protein, binds as a dimer to target DNA. EMBO J. 1987 Sep;6(9):2781–2784. doi: 10.1002/j.1460-2075.1987.tb02573.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kinnaird J. H., Keighren M. A., Kinsey J. A., Eaton M., Fincham J. R. Cloning of the am (glutamate dehydrogenase) gene of Neurospora crassa through the use of a synthetic DNA probe. Gene. 1982 Dec;20(3):387–396. doi: 10.1016/0378-1119(82)90207-4. [DOI] [PubMed] [Google Scholar]
  11. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  12. Mullis K. B., Faloona F. A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. doi: 10.1016/0076-6879(87)55023-6. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Paluh J. L., Orbach M. J., Legerton T. L., Yanofsky C. The cross-pathway control gene of Neurospora crassa, cpc-1, encodes a protein similar to GCN4 of yeast and the DNA-binding domain of the oncogene v-jun-encoded protein. Proc Natl Acad Sci U S A. 1988 Jun;85(11):3728–3732. doi: 10.1073/pnas.85.11.3728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Perkins D. D., Newmeyer D., Taylor C. W., Bennett D. C. New markers and map sequences in Neurospora crassa, with a description of mapping by duplication coverage, and of multiple translocation stocks for testing linkage. Genetica. 1969;40(3):247–278. doi: 10.1007/BF01787357. [DOI] [PubMed] [Google Scholar]
  16. Perkins D. D., Radford A., Newmeyer D., Björkman M. Chromosomal loci of Neurospora crassa. Microbiol Rev. 1982 Dec;46(4):426–570. doi: 10.1128/mr.46.4.426-570.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]

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