Chapter 16 : Cloning the Mating-Type Genes of : A Historical Perspective

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Preview this chapter:
Zoom in

Cloning the Mating-Type Genes of : A Historical Perspective, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555815837/9781555814212_Chap16-1.gif /docserver/preview/fulltext/10.1128/9781555815837/9781555814212_Chap16-2.gif


This chapter describes the life cycle of and ascribes function to the principal genetic determinants, the mating-type genes, that govern it, and presents the strategies that were devised for the successful isolation of these genes. In the world wide population of , a large number of A and B mating specificities are conferred by the extensive series of alleles of the genes residing at these loci. In and other Basidiomycetes, the genetic variability ensured by mating is further enhanced by the multiallelic mating-type system. Extraordinarily exact and regular in its supervision of mycelial interactions, the mating-type system of is not influenced by external factors such as nutrition and environmental conditions. Genomic DNA was isolated from a strain carrying the Aα4 allele to make the library used in the chromosomal walk from PAB1. The library was constructed in a cosmid vector in which the TRP1 gene would be the selectable marker in transformation. The mating genes are similar to other genes (and many fungal genes in general) in that there are no obvious promoter sequences such as a TATA box or CAAT box. Information from several previous studies was incorporated to create a successful strategy to isolate active fragments from the Bα and Bβ loci of .

Citation: Stankis M, Specht C. 2007. Cloning the Mating-Type Genes of : A Historical Perspective, p 267-282. In Heitman J, Kronstad J, Taylor J, Casselton L (ed), Sex in Fungi. ASM Press, Washington, DC. doi: 10.1128/9781555815837.ch16
Highlighted Text: Show | Hide
Loading full text...

Full text loading...


Image of Figure 16.1
Figure 16.1

(A) Life cycle of in diagrammatic representation. See the text for details. (B) Nuclear migration (shown for one mate). In a compatible interaction, a reciprocal exchange of nuclei takes place: in the fusion cell, the nucleus of each mate divides, and the daughter nuclei migrate into the cells of the other mating type. The donor nuclei rapidly divide, as they migrate throughout the existing mycelium of the other mate. Eventually they reach the apical cells. The nuclei move at a speed exceeding the rate of radial growth of the mycelia, i.e., at velocities of 1 to 6 mm per h, depending on the temperature ( ). At these velocities, a migrating nucleus can traverse one cell, which is typically 100 µm in length, in 1 min. Microtubules and microfibrils are associated with the migrating nuclei ( ). Nuclear migration requires the rapid dissolution of the septa between cells, a process that has been correlated with the production of a specific hydrolytic enzyme, R-glucanase ( ). Nuclear migration is regulated by the mating-type genes. (C) Conjugate nuclear division. The migrating nuclei arrive at the growing hyphal tips, where they pair, but do not fuse, with the resident nucleus. A mechanism for segregating paired nuclei, common to many Basidiomycetes, ensures that each newly formed cell has two nuclei, one from each parent. This is accomplished through the formation of a unique structure called a hook cell or clamp connection. Prior to nuclear division, a short branch arises on the side of the apical cell, into which one of the nuclei moves. Both nuclei then divide in synchrony and new cell walls form perpendicular to the spindles to generate three cells. One of each of the daughter nuclei is retained in the new apical cell, one in the hook cell and one in the subterminal cell. The hook or clamp cell then fuses with the subterminal cell, providing a bridge for the previously entrapped daughter nucleus to move into the subapical cell, where it pairs with its partner. This restores the dikaryotic condition to the penultimate cell. The presence of fused hook cells (clamp connections) can be used to identify dikaryotic mycelia. The formation of the hook cell and synchronized nuclear division are regulated by the genes, and the fusion of the hook cell with the subterminal cell is regulated by the genes. (D) Processes controlled by the and mating-type genes of . Adapted from Stankis et al. ( ).

Citation: Stankis M, Specht C. 2007. Cloning the Mating-Type Genes of : A Historical Perspective, p 267-282. In Heitman J, Kronstad J, Taylor J, Casselton L (ed), Sex in Fungi. ASM Press, Washington, DC. doi: 10.1128/9781555815837.ch16
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 16.2
Figure 16.2

(A) Selection strategy for the isolation of the mating-type gene. Following the isolation of the linked gene ( ), a chromosomal “walk” to 4 was accomplished by DNA-DNA hybridization of overlapping clones from a cosmid library ( ). Hypothesizing that the addition of 4 DNA to the genome of the recipient 1 strain, to create transformants merodiploid for , would activate -regulated developmental events, transformants at each step of the walk were screened by light microscopy for unfused hook cells. (B) Selection strategy for the isolation of the and mating-type genes. A mixture of protoplasts prepared from two strains having compatible mating types ( and ) and the same mating type (2 2) were transformed with a genomic library constructed with DNA from a b1 strain. Full sexual development, i.e., the formation of fruiting bodies, could occur if a transformant, merodiploid for either the or mating-type genes, mated with an -compatible regenerate, or fused with one during protoplast manipulation. Two fruiting transformants yielded haploid basidiospores which revealed the -activated phenotype ( ). The transforming and DNA was recovered using plasmid rescue techniques ( ).

Citation: Stankis M, Specht C. 2007. Cloning the Mating-Type Genes of : A Historical Perspective, p 267-282. In Heitman J, Kronstad J, Taylor J, Casselton L (ed), Sex in Fungi. ASM Press, Washington, DC. doi: 10.1128/9781555815837.ch16
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 16.3
Figure 16.3

(A) A generalized map of the mating-type region of Sequences demonstrating mating activity in , 3, 4, 5, and specificities average about 50% similarity, although they are embedded in a region of DNA common to all strains examined, as denoted by the shaded line. The transition from common sequence, encoding ( mitochondrial intermediate peptidase), at the left end, to the heterogeneous region is gradual in all mating types, except , where the transition (not shown) is extremely abrupt due to a natural deletion of the Z gene. The right boundary of the common sequence is approximately 7 to 8.5 kb from that of the left, except in , where heterogeneous DNA extends only 4.5 kb ( ). The and mating-type genes are a dyad of divergently transcribed homeodomain (HD1 and HD2) genes whose products regulate sexual development. Gene has no apparent function in mating. The coding region of each gene is boxed, and introns conserved among the alleles of each gene are shown as vertical black lines. The starred intron in HD2 is inserted between the codons for the highly conserved amino acid residues W and F. The direction of transcription is depicted by an arrow. GenBank accession numbers of DNA sequences used to generate map: , U13942 and M97179; , L43072, U13943, and M97180; , U13944 and M97181; 5, U22049; 6, AF274566. (B) A generalized map of the α Y and Z proteins and the β V6 protein. Each protein is depicted as a solid line with NH-and COOH-termini as denoted for α Y. The Y proteins all contain an HD2 homeodomain, a predicted coiled coil that overlaps a 28-residue bipartite nuclear localization sequence (NLS) that is embedded within a basic region ( ), and a serine-rich region (not shown). These characteristics have been observed for each Y protein studied to date, and the position of each feature is approximately the same in each Y allele. The Z proteins also contain features bearing on their presumptive function as transcriptional regulators, which reside in the same relative position in each allele: an atypical homeodomain sequence (HD1), two highly acidic 30-amino-acid regions rich in glutamate and aspartate (ARs), and two predicted coiled-coil regions (white ovals). The C-terminal regions of Z which are predicted to form a coiled coil display extremely high identity among the alleles ( ). The interacton of coiled coils is the mechanism by which both homo- and heterodimerizations of regulatory transcription factors occur in many organisms. Discussions and graphic presentations of the Y and Z regions which are involved in mating interactions, specificity, and binding are available ( ). The V6 protein, encoded by , displays an HD2 homeodomain motif. (C) The active regulatory complex encoded by and . Activation of development follows mating via the interaction of Y and Z proteins from compatible mates. An active complex is formed from the HD1 protein from one mate in combination with the HD2 protein from the other mate, that is, Yi in combination with Zj or Yj in combination with Zi. The heteromultimer is postulated to be a transcription factor that directs development down a new pathway by binding upstream of specific target genes ( ). The V protein, encoded at the locus, may be the HD2 partner of an HD1-HD2 pair, similar to the composition of the active products.

Citation: Stankis M, Specht C. 2007. Cloning the Mating-Type Genes of : A Historical Perspective, p 267-282. In Heitman J, Kronstad J, Taylor J, Casselton L (ed), Sex in Fungi. ASM Press, Washington, DC. doi: 10.1128/9781555815837.ch16
Permissions and Reprints Request Permissions
Download as Powerpoint


1. Asada, Y.,, C. Yue,, J. Wu,, G.-P. Shen,, C. Novotny, and, R. C. Ullrich. 1997. Schizophyllum commune Aα mating-type proteins, Y and Z, form complexes in all combinations in vitro. Genetics 147: 117123.
2. Asante-Owusu, R. N.,, A. H. Banham,, H. U. Böhnert, E. J. C. Mellor, and, L. A. Casselton. 1996. Heterodimerization between two classes of homeodomain proteins in the mushroom Coprinus cinereus brings together potential DNA-binding and activation domains. Gene 172: 2531.
3. Asgeirsdottir, S. A.,, F. H. J. Schuren, and, J. G. H. Wessels. 1994. Assignment of genes to pulse-field separated chromosomes of Schizophyllum commune. Mycol. Res. 98: 689693.
4. Beach, D. H. 1983. Cell type switching by DNA transposition in fission yeast. Nature 305: 682683.
5. Bertolino,, E., B. Reimund,, D. Wildt-Perinic, and R. G. Clerc. 1995. A novel homeobox protein which recognizes a TGT core and functionally interferes with a retinoid-responsive motif. J. Biol. Chem. 270: 3117831188.
6. Boulikas, T. 1994. Putative nuclear localization signals (NLS) in protein transcription factors. J. Cell. Biochem. 55: 3258.
7. Burglin,, T. R. 1997. Analysis of TALE superclass homeobox genes (MEIS, PBC, KNOX, Iroquios, TGIF) reveals a novel domain conserved between plants and animals. Nucleic Acids Res. 25: 41734180.
8. Burglin, T. R. 2003. The homeobox genes of Encephalitozoon cuniculi (Microsporidia) reveal a putative mating-type locus. Dev. Gene Evol. 213: 5052.
9. Burglin,, T. R. 2005. Homeodomain proteins, p. 179–222. In R. A. Meyers (ed.), Encyclopedia of Molecular Cell Biology and Molecular Medicine. Wiley-VCH Verlag GmbH & Co., Weinheim, Germany.
10. Dranginis, A. M. 1990. Binding of yeast a1 and alpha 2 as a heterodimer to the operator DNA of a haploid-specific gene. Nature (London) 347: 682685.
11. Fowler,, T. J., M. F. Mitton,, E. I. Rees, and, C. A. Raper. 2004. Crossing the boundary between the Bα and Bβ mating-type loci in Schizophyllum commune. Fungal Genet. Biol. 41: 89101.
12. Froeliger, E. H.,, A. Munoz-Rivas,, C. A. Specht,, R. C. Ullrich, and C. P. Novotny. 1987. The isolation of specific genes from the basidiomycete Schizophyllum commune. Curr. Genet. 12: 547554.
13. Giasson, L.,, C. A. Specht,, C. Milgrim,, C. P. Novotny, and, R. C. Ullrich. 1989. Cloning and comparison of Aα mating-type alleles of the Basidiomycete Schizophyllum commune. Mol. Gen. Genet. 218: 7277.
14. Gillissen, B.,, J. Bergemann,, C. Sandmann,, B. Schoee, and, M. Bolker, and, R. Kahmann. 1992. A two-component regulatory system for self/non-self recognition in Ustilago maydis. Cell 68: 647657.
15. Goutte, C., and, A. D. Johnson. 1988. a1 protein alters the DNA binding specificity of alpha 2 repressor. Cell 52: 875882.
16. Herskowitz, I. 1989. A regulatory hierarchy for cell specialization in yeast. Nature (London) 342: 749757.
17. Horton,, J. S., and C. A. Raper. 1991. Pulsed-field gel electrophoretic analysis of Schizophyllum commune chromosomal DNA. Curr. Genet. 19: 7780.
18. Isaya, G.,, W. R. Sakati,, R. A. Rollins,, G.-P. Shen,, L. C. Hanson,, R. C. Ullrich, and, C. P. Novotny. 1995. Mammalian mitochondrial intermediate peptidase: structure/ function analysis of a new homologue from Schizophyllum commune and relationship to thimet oligopeptidases. Genomics 28: 450461.
19. Johnson, A. D. 1995. Molecular mechanisms of cell-type determination in budding yeast. Curr. Opin. Genet. Dev. 5: 552558.
20. Koltin,, Y., J. R. Raper, and, G. Simchen. 1967. Genetic structure of the incompatibility factors of Schizophyllum commune: the B factor. Proc. Natl. Acad. Sci. USA 57: 5563.
21. Koltin, Y.,, J. Stamberg,, N. Bawnik,, R. Tamarkin, and, R. Werczberger. 1979. Mutational analysis of natural alleles in and affecting the B incompatibility factor of Schizophyllum. Genetics 93: 383391.
22. Kües, U., and, L. Casselton. 1992. Homeodomains and regulation of sexual development in basidiomycetes. Trends Genet. 8: 154155.
23. Kües, U.,, W. V. J. Richardson,, A. M. Tymon,, E. S. Mutasa,, B. Gottgens,, S. Gaubatz,, A. Gregoriades, and, L. A. Casselton. 1992. The combination of dissimilar alleles of the Aα and gene complexes, whose proteins contain homeodomain motifs, determines sexual development in the mushroom Coprinus cinereus. Genes Dev. 6: 568577.
24. Leonard, T. J., and, S. Dick. 1968. Chemical induction of haploid fruiting in Schizophyllum commune. Proc. Natl. Acad. Sci. USA 59: 745751.
25. Luo, Y.,, R. C. Ullrich, and, C. P. Novotny. 1994. Only one of the paired Schizophyllum commune Aα mating-type, putative homeobox genes encodes a homeodomain essential for Aa-regulated development. Mol. Gen. Genet. 244: 318324.
26. Magae, Y.,, C. Novotny, and, R. Ullrich. 1995. Interaction of the Aα Y and Z mating-type homeodomain proteins of Schizophyllum commune detected by the two-hybrid system. Biochem. Biophys. Res. Commun. 211: 10711076.
27. Marion, A. L.,, K. A. Bartholomew,, J. Wu,, H. Yang,, C. P. Novotny, and, R. C. Ullrich. 1995. The A mating-type locus of Schizophyllum commune: structure and function of gene X. Curr. Genet. 29: 143149.
28. Munoz-Rivas, A. M.,, C. A. Specht,, B. J. Drummond,, E. Froeliger,, C. P. Novotny, and, R. C. Ullrich. 1986. Transformation of the Basidiomycete Schizophyllum commune. Mol. Gen. Genet. 205: 103106.
29. Munoz-Rivas, A. M.,, C. A. Specht,, C. P. Novotny, and, R. C. Ullrich. 1986. Isolation of the DNA sequence coding indole-3-glycerol phosphate synthetase and phosphoribosylanthranilate isomerase of Schizophyllum commune. Curr. Genet. 10: 909913.
30. Niederpruem, D. J. 1980. Direct studies of dikaryotization in Schizophyllum commune. 1. Live intercellular nuclear migration patterns. Arch. Microbiol. 128: 162171.
31. Palmer,, G. E., and J. S. Horton. 2006. Mushrooms by magic: making connections between signal transduction and fruiting body development in the basidiomycete fungus Schizophyllum commune. FEMS Microbiol. Lett. 262: 18.
32. Parag, Y. 1962. Mutations in the B incompatibility factor of Schizophyllum commune. Proc. Natl. Acad. Sci. USA 48: 743750.
33. Raper,, C. A. 1978. Control of development by the incompatibility system in basidiomycetes, p. 3–29. In M. N. Schwalb and P. G. Miles (ed.), Genetics and Morphogenesis in the Basidiomycetes. Academic Press, New York, NY.
34. Raper, C. A. 1983. Controls for development and differentiation in the dikaryon in Basidiomycetes, p. 195–238. In J. Bennett and A. Ciegler (ed.), Secondary Metabolism and Differentiation in Fungi. Marcel Dekker, New York, NY.
35. Raper, C. A. 1988. Schizophyllum commune, a model for genetic studies of the Basidiomycotina, p. 511–522. In G. S. Sidhu (ed.), Genetics of Pathogenic Fungi, vol. 6 of D. S. Ingrains and, P. H. Williams (ed.), Advances in Plant Pathology. Academic Press, New York, NY.
36. Raper, C. A. 2004. Why study Schizophyllum? Fungal Genetics Newsl. 51: 3036.
37. Raper,, C.A., and J. R. Raper. 1966. Mutations modifying sexual morphogenesis in Schizophyllum. Genetics 54: 11511168.
38. Raper, C.A., and, J. R. Raper. 1973. Mutational analysis of a regulatory gene for morphogenesis in Schizophyllum. Proc. Natl. Acad. Sci. USA 70: 14271431.
39. Raper, J. R. 1966. Genetics of Sexuality in Higher Fungi. Ronald Press, New York, NY.
40. Raper, J. R.,, M. G. Baxter, and, A. H. Ellingboe. 1960. The genetic structure of the incompatibility factors of Schizophyllum commune: the A factor. Proc. Natl. Acad. Sci. USA 44: 889900.
41. Raudaskoski, M., and, R. Vauras. 1982. Scanning electron microscope study of fruit body differentiation in Schizophyllum commune. Trans. Br. Mycol. Soc. 78: 475481.
42. Raudaskoski, M.,, V. Salo, and, S. S. Niini. 1988. Structure and function of the cytoskeleton in filamentous fungi. Karstenia 28: 4960.
43. Robertson, C. I.,, K. A. Bartholomew,, C. P. Novotny, and, R. C. Ullrich. 1996. Deletion of the Schizophyllum commune Aα locus: the roles of Aα Y and Z mating-type genes. Genetics 144: 14371444.
44. Robertson, C. I.,, A. M. Kende,, K. Toenjes,, C. P. Novotny, and, R. C. Ullrich. 2002. Evidence for interaction of Schizophyllum commune Y mating-type proteins in vivo. Genetics 160: 14611467.
45. Ruiter, M. H.,, J. H. Sietsma, and, J. G. H. Wessels. 1988. Expression of dikaryon-specific mRNAs of Schizophyllum commune in relation to incompatibility genes, light, and fruiting. Exp. Mycol. 12: 6069.
46. Scott, M. P.,, J. W. Tamkun, and, G. W. Hartzell III. 1989. The structure and function of the homeodomain. Biochim. Biophys. Acta 989: 2548.
47. Shen, G.-P.,, Y. Chen,, D. Song,, Z. Peng,, C. P. Novotny, and, R. C. Ullrich. 2001. The Aα6 locus: its relation to mating-type regulation of sexual development in Schizophyllum commune. Curr. Genet. 39: 340345.
48. Shen, G.-P.,, D.-C. Park,, R. C. Ullrich, and, C. P. Novotny. 1996. Cloning and characterization of a Schizophyllum gene with Aβ6 mating-type activity. Curr. Genet. 29: 136142.
49. Specht, C. A. 1995. Isolation of the Bα and Bβ mating-types loci of Schizophyllum commune. Curr. Genet. 28: 374379.
50. Specht,, C. A., A. Munoz-Rivas,, C. P. Novotny, and, R. C. Ullrich. 1988. Transformation of Schizophyllum commune, an analysis of parameters for improving transformation frequencies. Exp. Mycol. 12: 357366.
51. Specht, C. A.,, M. M. Stankis,, L. Giasson,, C. P. Novotny, and, R. C. Ullrich. 1992. Functional analysis of the homeodomain-related proteins of the Aα locus of Schizophyllum commune. Proc. Natl. Acad. Sci. USA 89: 71747178.
52. Specht, C. A.,, M. M. Stankis,, C. P. Novotny, and, R. C. Ullrich. 1994. Mapping the heterogeneous DNA region that determines the nine Aα mating-type specificities of Schizophyllum commune. Genetics 137: 709714.
53. Stamberg, J., and, Y. Koltin. 1972. The organization of the incompatibility factors in higher fungi: the effects of structure and symmetry on breeding. Heredity 30: 1526.
54. Stankis, M. M.,, C. A. Specht, and, L. Giasson. 1990. Sexual incompatibility in Schizophyllum commune: from classical genetics to a molecular view. Semin. Dev. Biol. 1: 195206.
55. Stankis, M. M.,, C. A. Specht,, H. Yang,, L. Giasson,, R. C. Ullrich, and, C. P. Novotny. 1992. The Aα mating locus of Schizophyllum commune encodes two dissimilar multiallelic homeodomain proteins. Proc. Natl. Acad. Sci. USA 89: 71697173.
56. Strathern, J. N.,, E. Spatola,, C. McGill, and J. B. Hicks. 1980. Structure and organization of transposable mating-type cassettes in Saccharomyces yeast. Proc. Natl. Acad. Sci. USA 77: 28392843.
57. Ullrich, R. C.,, L. Giasson,, C. A. Specht,, M. M. Stankis, and, C. P. Novotny. 1990. The Aα multiallelic mating-type genes of Schizophyllum commune, p. 271–288. In E. H. Davidson,, J. V. Ruderman, and, J. W. Posakony (ed.), Developmental Biology. Wiley-Liss, New York, NY.
58. Vaillancourt, L. J.,, M. Raudaskoski,, C. A. Specht, and, C. A. Raper. 1997. Multiple genes encoding pheromones and a pheromone receptor define the Bβ1 mating-type specificity in Schizophyllum commune. Genetics 146: 541551.
59. Wendland, J.,, L. J. Vaillancourt,, J. Hegner,, K. B. Lengeler,, K. J. Laddison,, C. A. Specht,, C. A. Raper, and, E. Kothe. 1995. The mating-type locus Bβ1 of Schizophyllum commune contains a pheromone receptor gene and putative pheromone genes. EMBO J. 14: 52715278.
60. Wessels, J. G. H. 1992. Gene expression during fruiting in Schizophyllum commune. Mycol. Res. 96: 609620.
61. Wessels,, J. G. H., and J. R. Marchant. 1974. Enzymatic degradation in hyphal wall preparations from a monokaryon and a dikaryon of Schizophyllum commune. J. Gen. Microbiol. 83: 359368.
62. Wolberger, C. 1999. Multiprotein-DNA complexes in transcriptional regulation. Annu. Rev. Biophys. Biomol. Struct. 28: 2956.
63. Wolberger,, C., A. K. Vershon,, B. Liu,, A. D. Johnson, and, C. O. Pabo. 1991. Crystal structure of a MAT alpha 2 homeodomain-operator complex suggests a general model for homeodomain-DNA interactions. Cell 67: 517528.
64. Wu, J.,, R. C. Ullrich, and, C. P. Novotny. 1996. Regions in the Z5 mating gene of Schizophyllum commune involved in Y-Z binding and recognition. Mol. Gen. Genet. 252: 739745.
65. Yang, H.,, G.-P. Shen,, D. C. Park,, C. P. Novotny, and, R. C. Ullrich. 1995. The Aα mating-type transcripts of Schizophyllum commune. Exp. Mycol. 19: 1625.
66. Yue, C.,, M. Osier,, C. P. Novotny, and, R. C. Ullrich. 1997. The specificity determinant of the Y mating-type proteins of Schizophyllum commune is also essential for Y-Z protein binding. Genetics 145: 253260.

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error