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The molecular nature of mutations in themt A-1 gene of theNeurospora crassa A idiomorph and their relation to mating-type function

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Abstract

The 293-amino acidmt A-1 ORF of theA mating-type idiomorph ofNeurospora crassa is multifunctional. It confersA mating identity and is responsible for heterokaryon incompatibility. The goal of this study was to dissect the functional regions ofmt A-1. New mutants ofmt A-1 selected for loss of the incompatibility function were obtained. One new mutant,A m99, was partially fertile as a maternal parent. This is the first time that fertility and incompatibility functions have been separated for theA idiomorph. In this mutant, themt A-1 ORF is truncated after the first 85 amino acids, indicating that this N-terminal region is minimally sufficient for female fertility. A series of deletion constructs and frameshift alleles ofmt A-1 was obtained and tested for male-mating activity and vegetative incompatibility in transformation experiments. These experiments showed that a region from position 1 to 111 is sufficient to confer incompatibility, while amino acids from position 1 to 227 are required for mating activity. A transcriptional analysis ofmt A-1 showed that the mRNA is expressed both before and after fertilization. This, together with the phenotype of theA m99 mutant, suggests a post-fertilization function formt A-1.

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References

  • Arnaise S, Zickler D, Glass NL (1993) Heterologous expression of mating-type genes in filamentous fungi. Proc Natl Acad Sci USA 90:6616–6620

    PubMed  Google Scholar 

  • Astell CR, Ahlstrom-Jonasson L, Smith M, Tatchell K, Nasmyth KA, Hall BD (1981) The sequence of the DNAs coding for the mating-type loci ofSaccharomyces cerevisiae. Cell 27:15–23

    PubMed  Google Scholar 

  • Beadle GW, Coonradt VL (1944) Heterokaryosis inNeurospora crassa. Genetics 29:291–308

    Google Scholar 

  • Bistis GN (1983) Evidence for diffusible, mating-type-specific trichogyne attractants inNeurospora crassa. Exp Mycol 7:292–295

    Google Scholar 

  • Brockman HE, Deserres FJ, Ong TM, Hung CY (1987) Two N-hydroxylaminopurines are highly mutagenic in thead-3 forward-mutation test in growing cultures of heterokaryon 12 ofNeurospora crassa. Mutat Res 177:61–75

    PubMed  Google Scholar 

  • Bruchez JJP, Eberle J, Russo VEA (1993) Regulatory sequences in the transcription ofNeurospora crassa genes: CAAT box, TATA box, introns, poly(A) tail formation sequences. Fungal Genet Newslett 40:89–96

    Google Scholar 

  • Carrol AM, Sweigard JA, Valent B (1994) Improved vectors for selecting resistance to hygromycin. Fungal Genet Newslett 41:22

    Google Scholar 

  • Casselton LA (1993) Fungal mating type genes — regulators of sexual development. Mycol Res 96:993–1006

    Google Scholar 

  • Chang S, Staben C (1994) Directed replacement ofmt-A bymt a-1 effects a mating-type switch inNeurospora crassa. Genetics 138:75–81

    PubMed  Google Scholar 

  • Debuchy R, Coppin E (1992) The mating types ofPodospora anserina: functional analysis and sequence of the fertilization domains. Mol Gen Genet 233:113–121

    PubMed  Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395

    PubMed  Google Scholar 

  • Dodge BO (1935) The mechanics of sexual reproduction inNeurospora. Mycologia 27:418–436

    Google Scholar 

  • Edelman S, Staben C (1994) A statistical analysis of sequence features within genes fromNeurospora crassa. Exp Mycol 18:70–81

    Google Scholar 

  • Fourney RM, Miyakoshi J, Day RS, Paterson MC (1988) Northern blotting: efficient RNA staining and transfer. Focus 10:5–7

    Google Scholar 

  • Frohman MA, Dush MK, Martin GR (1988) Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci USA 85:8998–9002

    PubMed  Google Scholar 

  • Glass NL, Kuldau GA (1992) Mating type and vegetative incompatibility in filamentous ascomycetes. Annu Rev Phytopathol 30:201–224

    Google Scholar 

  • Glass NL, Lee L (1992) Isolation ofNeurospora crassa A mating type mutants by repeat induced point mutation. Genetics 132:125–133

    PubMed  Google Scholar 

  • Glass NL, Nelson MA (1994) Mating-type genes in mycelial ascomycetes. In: Wessels JDH, Meidhardt F (eds) The Mycota: growth, differentiation and sexuality. Springer, Berlin New York Heidelberg pp 295–306

    Google Scholar 

  • Glass NL, Staben C (1990) Genetic control of mating inNeurospora crassa. Semin Dev Biol 1:177–184

    Google Scholar 

  • Glass NL, Vollmer SJ, Staben C, Grotelueschen J, Metzenberg RL, Yanofsky C (1988) DNAs of the two mating-type alleles ofNeurospora crassa are highly dissimilar. Science 241:570–573

    PubMed  Google Scholar 

  • Glass NL, Grotelueschen J, Metzenberg RL (1990)Neurospora crassa A mating-type region. Proc Natl Acad Sci USA 87:4912–4916

    PubMed  Google Scholar 

  • Griffiths AFJ (1982) Null mutants of theA and a mating-type alleles ofNeurospora crassa. Can J Genet Cytol 24:167–176

    Google Scholar 

  • Griffiths AJF, Delange AM (1978) Mutations of the a mating type inNeurospora crassa. Genetics 88:239–254

    Google Scholar 

  • Hagen DC, Bruhn L, Westby CA, Sprague GF Jr (1993) Transcription of a-specific genes inSaccharomyces cerevisiae: DNA sequence requirements for activity of the coregulatorα1. Mol Cell Biol 13:6866–6875

    PubMed  Google Scholar 

  • Herskowitz I (1988) Life cycle of the budding yeastSaccharomyces cerevisiae. Microbiol Rev 52:536–553

    PubMed  Google Scholar 

  • Logemann J, Schell J, Willmitzer L (1987) Improved method for isolation of RNA from plant tissues. Anal Biochem 163:16–20

    PubMed  Google Scholar 

  • Metzenberg RL, Glass NL (1990) Mating type and mating strategies inNeurospora. Bio essays 12:53–59

    Google Scholar 

  • Newmeyer D (1970) A suppressor of the heterokaryon-incompatibility associated with mating type inNeurospora crassa. Can J Genet Cytol 12:914–926

    PubMed  Google Scholar 

  • Philley ML, Staben C (1994) Functional analysis of theNeurospora crassa MT a-1 mating type polypeptide. Genetics 137:715–722

    PubMed  Google Scholar 

  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Ehrlich HA (1988) Primer directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491

    PubMed  Google Scholar 

  • Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    PubMed  Google Scholar 

  • Schweizer M, Case ME, Dykstra CC, Giles NH, Kushner SR (1981) Identification and characterization of recombinant plasmids carrying the completeqa gene cluster fromNeurospora crassa including theqa-1 + regulatory gene. Proc Natl Acad Sci USA 78:5086–5090

    PubMed  Google Scholar 

  • Staben C, Yanofsky C (1990)Neurospora crassa a mating-type region. Proc Natl Acad Sci USA 87:4917–4921

    PubMed  Google Scholar 

  • Stevens JN, Metzenberg RL (1982) Preparing Neurospora DNA: some improvements. Neurospora Newslett 29:27–28

    Google Scholar 

  • Tan S, Richmond TJ (1990) DNA binding-induced conformational change of the yeast transcriptional activator PRTF. Cell 62:367–377

    PubMed  Google Scholar 

  • Tan S, Ammerer G, Richmond TJ (1988) Interactions of purified transcription factors: binding of yeast MATα1 and PRTF to cell-type specific, upstream activating sequences. EMBO J 7:4255–4264

    PubMed  Google Scholar 

  • Turgeon BG, Bohlmann H, Ciuffetti LM, Christiansen SK, Yang G, Schafer W, Yoder OC (1993) Cloning and analysis of the mating type genes fromCochhobolus heterostrophus. Mol Gen Genet 238:270–284

    PubMed  Google Scholar 

  • Vellani TS, Griffiths AJF, Glass NL (1994) New mutations that suppress mating-type vegetative incompatibility inNeurospora crassa. Genome 37:249–255

    PubMed  Google Scholar 

  • Vogel HJ (1964) Distribution of lysine pathways among fungi: evolutionary implications. Am Nat 98:435–466

    Google Scholar 

  • Vollmer SJ, Yanofsky C (1986) Efficient cloning of genes ofNeurospora crassa. Proc Natl Acad Sci USA 83:4869–4873

    Google Scholar 

  • Westergaard M, Mitchell HK (1947)Neurospora V. A synthetic medium favoring sexual reproduction. Am J Bot 34:573–577

    Google Scholar 

  • Yuan YO, Stroke IL, Fields S (1993) Coupling of cell identity to signal response in yeast: interaction between theα1 and STE12 proteins. Genes Dev 7:1584–1597

    PubMed  Google Scholar 

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Author notes

  1. Leisa Stenberg

    Present address: Department of Biochemistry and Molecular Biology, University of British Columbia, V6T 1Z3, Vancouver, B.C., Canada

Authors and Affiliations

  1. Department of Botany, University of British Columbia, 6270 University Boulerard, V6T 1Z4, Vancouver, B.C., Canada

    Sven Saupe, Leisa Stenberg, Ka Tai Shiu, Anthony J. F. Grifiths & N. Louise Glass

  2. Biotechnology Laboratory, University of British Columbia, V6T 1Z3, Vancouver, B.C., Canada

    Sven Saupe, Ka Tai Shiu, Anthony J. F. Grifiths & N. Louise Glass

Authors
  1. Sven Saupe
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  2. Leisa Stenberg
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  3. Ka Tai Shiu
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  4. Anthony J. F. Grifiths
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  5. N. Louise Glass
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Communicated by C. A. M. J. J. Van den Hondel

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Saupe, S., Stenberg, L., Shiu, K.T. et al. The molecular nature of mutations in themt A-1 gene of theNeurospora crassa A idiomorph and their relation to mating-type function. Molec. Gen. Genet. 250, 115–122 (1996). https://doi.org/10.1007/BF02191831

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  • DOI: https://doi.org/10.1007/BF02191831

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