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  • Homeodomain  (3)
  • Gene isolation  (2)
  • Key words Schizophyllum commune  (1)
  • 1
    ISSN: 1432-0983
    Keywords: Key words Schizophyllum commune ; Mating type ; Homeodomain
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The A-pathway of development in the basidiomycete fungus Schizophyllum commune may be activated by either of two mating-type loci, Aα and Aβ. Aα consists of two multiallelic genes, Y and Z. Y contains a putative homeodomain; Z contains a homeodomain-related region. Non-self combinations of Y and Z form heteromultimers which are thought to be transcription factors of developmental genes. To more completely understand A-regulated development it is necessary to address the issue of functional redundancy, i.e., how do two different mating loci, Aα and Aβ, both manage to regulate the same pathway. Here we report the structure of a gene with Aβ6 activity. This gene, denoted AβV6, encodes a deduced polypeptide of 640 amino-acids with a homeodomain motif. V6 also contains a 20-amino acid sequence that is conserved in Aα Y1, Y3 and Y4. Except for the homeodomain and the conserved sequence, the deduced V6 polypeptide shows no significant identity to AαY, AαZ, or other known proteins. The presence of a homeodomain suggests that V, like Y and Z, may be a regulatory protein for genes in the A-pathway. Thus while Aα and Aβ encode different proteins, the general mechanism by which Aα and Aβ components signal A-regulated development may be similar.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0983
    Keywords: E. coli trpC ; Schizophyllum TRP1 ; Complementation ; Gene isolation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A Schizophyllum gene library was made in plasmid pRK9, Plasmids from this library were tested for their ability to complement several auxotrophic mutations of Escherichia coli. The goal was to isolate a Schizophyllum auxotrophic gene that could be used to transform a corresponding Schizophyllum auxotrophic mutant to prototrophy. Complementation was observed only for E. coli trpC indole 3-glycerol phosphate synthetase (IGPS) and phosphoribosyl-anthranilate isomerase (PRAI) mutations. Plasmids with a Schizophyllum sequence coding for both IGPS and PRAI activities were recovered from E. coli transformants. Expression of the Schizophyllum gene (TRP1) in E. coli is probably dependent on the Serratia marcescens promoter of plasmid pRK9. The DNA sequence containing the Schizophyllum TRP1 gene was not obviously rearranged in cloning.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0983
    Keywords: Schizophyllum commune ; Transformation ; Gene isolation ; Basidiomycetes ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have developed a routine way to isolate genes directly from the basidiomycete fungus, Schizophyllum commune. Plasmid DNA from a genomic gene library was used to isolate five specific genes by complementation of Schizophyllum mutations via transformation. The mutant strains were deficient in the ability to synthesize either adenine (ade2 and ade5), uracil (ural, encoding orotidine-5′-phosphate decarboxylase; OMPdecase), tryptophan (rpl, encoding indole-3-glycerol phosphate synthetase; IGPS) or para aminobenzoic acid (pab1). In each case, Southern analysis revealed that transformation to prototrophy was concomitant with the integration of vector sequence into the genome of the S. commune mutant. Total DNA from transformants was restricted, religated, and used to transform E. coli. Ampicillin resistant plasmids were recovered from E. coli and tested for their ability to transform the corresponding mutant of S. commune. Plasmids complementing the ade2, adeS, pabl, trpl, and ural mutations were recovered.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0983
    Keywords: Schizophyllum commune ; Mating type ; Homeodomain
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The A-pathway of development in the basidiomycete fungusSchizophyllum commune may be activated by either of two mating-type loci,Aξ andAβ Aα consists of two multiallelic genes,Y andZ. Y contains a putative homeodomain; Z contains a homeodomain-related region. Non- self combinations of Y and Z form heteromultimers which are thought to be transcription factors of developmental genes. To more completely understand A-regulated development it is necessary to address the issue of functional redundancy, i.e., how do two different mating loci,Aα andAβ, both manage to regulate the same pathway. Here we report the structure of a gene withAβ6 activity. This gene, denotedAβV6, encodes a deduced polypeptide of 640 amino-acids with a homeodomain motif. V6 also contains a 20-amino acid sequence that is conserved in Aα Y1, Y3 and Y4. Except for the homeodomain and the conserved sequence, the deduced V6 polypeptide shows no significant identity to AαY, AαZ, or other known proteins. The presence of a homeodomain suggests that V, like Y and Z, may be a regulatory protein for genes in the A-pathway. Thus whileAα andAβ encode different proteins, the general mechanism by which Aα and Aβ components signal A-regulated development may be similar.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1617-4623
    Keywords: Schizophyllum commune ; Homeodomain ; Homeobox ; Mating type
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Aα mating-type locus is one of four master regulatory loci controlling sexual development in Schizophyllum commune. The Aα locus contains two homeobox genes, Y and Z, encoding two homeodomain-related proteins, Y and Z, Y and Z are each multi-allelic genes. When haploid strains form fusion cells, only particular combinations of Y and Z alleles activate Aα-regulated sexual development. The role of the putative homeodomain was examined in several Y and Z alleles by site-directed mutagenesis of regions critical to secondary structure and function of homeodomains. Mutations of the Z homeobox do not affect the function of Z proteins in Aα-activated development, but mutations of Yhomeoboxes destroy the ability of Y proteins to activate development. We conclude that only one of two Aα homeodomain-related regulators relies upon the homeodomain motif to effect gene expression in sexual development. This conclusion affords a refinement of our working hypothesis for the mechanism by which Aα proteins may regulate target gene expression. On the basis of our results with the Z protein, we speculate that the DNA-binding motifs of some transcriptional regulators may be lost or modified during evolution once these regulators have been recruited to participate in complexes with other DNA-binding proteins.
    Type of Medium: Electronic Resource
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