Abstract
Cochliobolus heterostrophus, a heterothallic Ascomycete, has a single mating type locus with two alternate forms called MAT-1 and MAT-2. MAT-1 was cloned by complementing a MAT-2 strain using a cosmid library from a MAT-1 strain and screening for a homothallic transformant. The cosmid recovered from this transformant was able to re-transform a MAT-2 strain to homothallism and MAT identity was proven by restriction fragment length polymorphism and conventional genetic mapping. All homothallic transformants could mate with either MAT-1 or MAT-2 strains, although the number of ascospores produced by self matings or crosses to MAT-2 strains was low. Progeny of selfed homothallic transformants were themselves homothallic. MAT-2 was cloned by probing a cosmid library from a MAT-2 strain with a fragment of insert DNA from the MAT-1 cosmid. A 1.5 kb subclone of either MAT-containing cosmid was sufficient to confer mating function in transformants. Examination of the DNA sequence of these subclones revealed that MAT-1 and MAT-2 contain 1297 by and 1171 bp, respectively, of completely dissimilar DNA flanked by DNA common to both mating types. Putative introns were found (one in each MAT gene) which, when spliced out, would yield open reading frames (ORFs) that occupied approximately 90% of the dissimilar DNA sequences. Translation of the MAT-1 ORF revealed similarity to the Neurospora crassa MATA, Podospora anserina mat−, and Saccharomyces cerevisiae MATα1 proteins; translation of the MAT-2 ORF revealed similarity to the N. crassa MATa, P. anserina mat+, and Schizosaccharomyces pombe mat-Mc proteins. These gene products are all proven or proposed DNA binding proteins. Those with similarity to MAT-2 are members of the high mobility group.
Similar content being viewed by others
References
Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410
Astell C, Ahlstrom-Jonasson L, Smith M, Tatchell K, Nasmyth K, Hall B (1981) The sequence of the DNAs coding for the mating-type loci of Saccharomyces cerevisiae. Cell 27:15–23
Ballance DJ (1990) Transformation systems for filamentous fungi and an overview of fungal gene structure. In: Leong SA, Berka RM (eds) Molecular industrial mycology: systems and applications for filamentous fungi. Marcel Dekker, New York, pp 1–29
Debuchy R, Coppin E (1992) The mating types of Podospora anserina: Functional analysis and sequence of the fertilization domains. Mol Gen Genet 233:113–121
Egel R, Neilson O, Weilguny D (1990) Sexual differentiation in fission yeast. Trends Genet 6:369–373
Feinberg AP, Vogelstein B (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13
Giasson L, Specht CA, Milgrim C, Novotny CP, Ullrich RC (1989) Cloning and comparison of Aa mating-type alleles of the Basidiomycete Schizophyllum commune. Mol Gen Genet 218:72–77
Glass NL, Vollmer SJ, Staben C, Grotelueschen J, Metzenberg RL, Yanofsky C (1988) DNAs of the two mating-type alleles of Neurospora crassa are highly dissimilar. Science 241:570–573
Glass NL, Grotelueschen J, Metzenberg RL (1990a) Neurospora crassa A mating-type region. Proc Natl Acad Sci USA 87:4912–4916
Glass NL, Metzenberg RL, Raju NB (1990b) Homothallic Sordariaceae from nature: the absence of strains containing only the a mating type sequence. Exp Mycol 14:274–289
Guzman D, Garber RC, Yoder OC (1982) Cytology of meiosis I and chromosome number of Cochliobolus heterostrophus (Ascomycetes). Can J Bot 60:1138–1141
Henikoff S (1984) Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351–359
Herskowitz I (1988) The life cycle of the budding yeast Saccharomyces cerevisiae. Microbiol Rev 52:536–553
Herskowitz I (1989) A regulatory hierarchy for cell specialization in yeast. Nature 342:749–757
Hicks J, Strathern JN, Klar AJS (1979) Transposable mating type genes in Saccharomyces cerevisiae. Nature 282:478–483
Holmes DS, Quigley M (1981) A rapid boiloing method for the preparation of bacterial plasmids. Anal Biochem 114:193–197
Jantzen HM, Admon A, Bell SP, Tjian R (1990) Nucleolar transcription factor hUBF contains a DNA binding motif with homology to HMG proteins. Nature 344:830–836
Kelly M, Burke J, Smith M, Klar A, Beach D (1988) Four mating-type genes control sexual differentiation in the fission yeast. EMBO J 7:1537–1548
Klar AJS, Strathern JN, Broach JR, Hicks JB (1981) Regulation of transcription in expressed and unexpressed mating type cassettes of yeast. Nature 289:239–252
Kronstad JW, Leong SA (1989) Isolation of two alleles of the b locus of Ustilago maydis. Proc Natl Acad Sci USA 86:978–982
Leach J, Yoder OC (1983) Heterokaryon incompatibility in the plant pathogenic fungus Cochliobolus heterostrophus. J Hered 74:149–152
Leach J, Lang BR, Yoder OC (1982) Methods for selection of mutants and in vitro culture of Cochliobolus heterostrophus. J Gen Microbiol 128:1719–1729
Mackay V, Manney TR (1974) Mutations affecting sexual conjugation and related processes in Saccharomyces cerevisiae. I. Isolation and phenotypic characterization of nonmating mutants. Genetics 76:255–271
Marsh L, Neiman AM, Herskowitz I (1991) Signal transduction during pheromone response in yeast. Annu Rev Cell Biol 7:699–728
Metzenberg RL (1990) The role of similarity and difference in fungal mating. Genetics 125:457–462
Metzenberg RL, Glass NL (1990) Mating type and mating strategies in Neurospora. Bioessays 12:53–59
Meyerowitz EM, Guild GM, Prestidge LS, Hogness DS (1980) A new high capacity cosmid vector and its use. Gene 11:271–282
Miller AM (1984) The yeast MATa1 gene contains two introns. EMBO J 3:1061–1065
Mutasa ES, Tymon AM, Gottgens B, Mellon FM, Little PFR, Casselton LA (1990) Molecular organization of an A mating type factor of the basidiomycete fungus Coprinus cinereus. Curr Genet 18:223–229
Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448
Picard M, Debuchy R, Coppin E (1991) Cloning the mating types of the heterothallic fungus Podospora anserina — Developmental features of haploid transformants carrying both mating types. Genetics 128:539–547
Raju NB (1980) Meiosis and ascospore genesis in Neurospora. Eur J Cell Biol 23:208–223
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Sinclair AH, Berta P, Palmer MS, Hawkins JR, Griffiths BL, Matthijs JS, Foster JW, Frischauf AM, LovellBadge R, Goodfellow PM (1990) A gene from the human sex-determining region encodes a protein with homology to a conserved DNA binding motif. Nature 346:240–244
Staben C, Yanofsky C (1990) Neurospora crassa a mating type region. Proc Natl Acad Sci USA 87:4917–4921
Tartof KD, Hobbs CA (1987) Improved media for growing plasmid and cosmid clones. Focus 9:12
Tatchell K, Nasmyth KA, Hall BD, Astell C, Smith M (1981) In vitro mutation analysis of the mating-type locus in yeast. Cell 27:25–35
Tsuda KI, Kikuchi M, Mori K, Waga S, Yoshida M (1988) Primary structure of non-histone protein HMGl revealed by the nucleotide sequence. Biochemistry 27:6159–6163
Turgeon BG, Garber RC, Yoder OC (1985) Transformation of the fungal maize pathogen Cochliobolus heterostrophus using the Aspergillus nidulans amdS gene. Mol Gen Genet 201:450–453
Turgeon BG, Garber RC, Yoder OC (1987) Development of a fungal transformation system based on selection of sequences with promoter activity. Mol Cell Biol 7:3297–3305
Turgeon BG, Ciuffetti L, Schafer W, Yoder OC (1988) Isolation of the mating type locus of Cochliobolus heterostrophus. In: Palacios R, Verma DPS (eds) Molecular Genetics of Plant-Microbe Interactions. APS Press, St Paul, pp 265–266
Wagstaff JE, Klapholz S, Easton-Esposito RE (1982) Meiosis in haploid yeast. Proc Natl Acad Sci USA 79:2986–2990
Wright JM, Dixon GH (1988) Induction by torsional stress of an altered DNA conformation 5′ upstream of the gene for a high mobility group protein from trout and specific to flanking sequences by the gene product HMG-T. Biochemistry 27:576–581
Yelton MM, Timberlake WE, van den Hondel CAMJJ (1985) A cosmid for selecting genes by complementation in Aspergillus nidulans: Selection of the developmentally regulated yA locus. Proc Natl Acad Sci USA 82:834–838
Yoder OC (1988) Cochliobolus heterostrophus, cause of Southern Corn Leaf Blight. In: Sidhu GS (ed) Genetics of plant pathogenic fungi. Academic Press, San Diego, pp 93–112
Yoder OC, Valent B, Chumley F (1986) Genetic nomenclature and practice for plant pathogenic fungi. Phytopathology 76:383–385
Yoder OC, Turgeon BG, Schäfer W, Ciuffetti LM, Bohlmann H, VanEtten HD (1989) Molecular analysis of mating type and expression of a foreign pathogenicity gene in Cochliobolus heterostrophus. In: Nevalainen H, Pentilla M (eds) Molecular biology of filamentous fungi. Foundation for Biotechnical and Industrial Fermentation Research, Helsinki, pp 189–196
Author information
Authors and Affiliations
Additional information
Communicated by C.A.M.J.J. van den Hondel
The first three authors contributed equally to the work
Rights and permissions
About this article
Cite this article
Gillian Turgeon, B., Bohlmann, H., Ciuffetti, L.M. et al. Cloning and analysis of the mating type genes from Cochliobolus heterostrophus . Molec. Gen. Genet. 238, 270–284 (1993). https://doi.org/10.1007/BF00279556
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00279556