scholarly journals Multiple Functions of mfa-1, a Putative Pheromone Precursor Gene of Neurospora crassa

2002 ◽  
Vol 1 (6) ◽  
pp. 987-999 ◽  
Author(s):  
Hyojeong Kim ◽  
Robert L. Metzenberg ◽  
Mary Anne Nelson
Keyword(s):  
Neurospora Crassa ◽  
Sexual Development ◽  
Vegetative Growth ◽  
Cysteine Residue ◽  
Northern Analysis ◽  
Mating Types ◽  
Wild Type ◽  
Minute Amount ◽  
Female Sexual Development ◽  
Ascospore Production

ABSTRACT A putative pheromone precursor gene of Neurospora crassa, mfa-1 (which encodes mating factor a-1), was identified as the most abundant clone in starved mycelial and perithecial cDNA libraries. Northern analysis demonstrated high mfa-1 expression in all mating type a tissues and suggested low expression levels in mat A tissues. The mfa-1 gene was expressed as an approximately 1.2-kb transcript predicted to encode a 24-residue peptide, followed by a long 3′ untranslated region (3′ UTR). The predicted MFA1 sequence showed 100% sequence identity to PPG2 of Sordaria macrospora and structural similarity (a carboxy-terminal CAAX motif) to many hydrophobic fungal pheromone precursors. Mutants with a disrupted open reading frame (ORF) in which the critical cysteine residue had been changed to a nonprenylatable residue, tyrosine (YAAX mutants), were isolated, as were mfa-1 mutants with intact ORFs but multiple mutations in the 3′ noncoding region (CAAX mutants). The 3′ UTR is required for the full range of mfa-1 gene activity. Both classes of mutants showed delayed and reduced vegetative growth (which was suppressed by supplementation with a minute amount [30 μM] of ornithine, citrulline, or arginine), as well as aberrant sexual development. When crossed as female parents to wild-type males, the CAAX and YAAX mutants showed greatly reduced ascospore production. No ascospores were produced in homozygous mfa-1 crosses. As males, YAAX mat a mutants were unable to attract wild-type mat A trichogynes (female-specific hyphae) or to initiate sexual development, while CAAX mat a mutants were able to mate and produce sexual progeny despite their inability to attract mat A trichogynes. In the mat A background, both CAAX and YAAX mutants showed normal male fertility but defective vegetative growth and aberrant female sexual development. Thus, the mfa-1 gene appears to have multiple roles in N. crassa development: (i) it encodes a hydrophobic pheromone with a putative farnesylated and carboxymethylated C-terminal cysteine residue, required by mat a to attract trichogynes of mat A; (ii) it is involved in female sexual development and ascospore production in both mating types; and (iii) it functions in vegetative growth of both mating types.

Characterization of mat A-2, mat A-3 and ΔmatA Mating-Type Mutants of Neurospora crassa

Genetics ◽  
1998 ◽  
Vol 148 (3) ◽  
pp. 1069-1079 ◽  
Author(s):  
Adlane V-B Ferreira ◽  
Zhiqiang An ◽  
Robert L Metzenberg ◽  
N Louise Glass
Keyword(s):  
Neurospora Crassa ◽  
Mating Type ◽  
Sexual Development ◽  
Vegetative Growth ◽  
Double Mutant ◽  
Sexual Cycle ◽  
Wild Type ◽  
Type Locus ◽  
Isolation And Characterization

AbstractThe mating-type locus of Neurospora crassa regulates mating identity and entry into the sexual cycle. The mat A idiomorph encodes three genes, mat A-1, mat A-2, and mat A-3. Mutations in mat A-1 result in strains that have lost mating identity and vegetative incompatibility with mat a strains. A strain containing mutations in both mat A-2 and mat A-3 is able to mate, but forms few ascospores. In this study, we describe the isolation and characterization of a mutant deleted for mat (ΔmatA), as well as mutants in either mat A-2 or mat A-3. The ΔmatA strain is morphologically wild type during vegetative growth, but it is sterile and heterokaryon compatible with both mat A and mat a strains. The mat A-2 and mat A-3 mutants are also normal during vegetative growth, mate as a mat A strain, and produce abundant biparental asci in crosses with mat a, and are thus indistinguishable from a wild-type mat A strain. These data and the fact that the mat A-2 mat A-3 double mutant makes few asci with ascospores indicate that MAT A-2 and MAT A-3 are redundant and may function in the same pathway. Analysis of the expression of two genes (sdv-1 and sdv-4) in the various mat mutants suggests that the mat A polypeptides function in concert to regulate the expression of some sexual development genes.

A SUPPRESSOR OF THE HETEROKARYON- INCOMPATIBILITY ASSOCIATED WITH MATING TYPE IN NEUROSPORA CRASSA

1970 ◽  
Vol 12 (4) ◽  
pp. 914-926 ◽  
Author(s):  
Dorothy Newmeyer
Keyword(s):  
Linkage Group ◽  
Neurospora Crassa ◽  
Mating Type ◽  
Vegetative Growth ◽  
Mode Of Action ◽  
Wild Strain ◽  
Group Iv ◽  
Mating Types ◽  
Vegetative Incompatibility ◽  
Opposite Mating Type

Neurospora crassa strains of opposite mating type are ordinarily heterokaryon-incompatible during vegetative growth. An unlinked mutant called tolerant (tol) is described, which suppresses the vegetative incompatibility of unlike mating types without affecting their ability to cross. The mutant tol was selected and studied by means of duplications heterozygous for mating type. Use of the duplication eliminates complications due to unlinked heterokaryon genes. The mode of action of tol has been confirmed by conventional heterokaryon tests. tol has been mapped in linkage group IV, close to tryp-4. A suppressor similar or identical to tolerant has been found in a wild strain from Panama, out of 14 different wild types which were tested. By using a different duplication which covers the unlinked heterokaryon-compatibility locus C, it was shown that tolerant does not suppress C/c incompatibility. The fact that tolerant suppresses only one of the two functions ascribed to mating type revives the question of whether 'mating-type' is one gene or two. However, the data strongly support Pittenger's (1957) conclusion that, if two genes are involved, they must be closely linked.

scholarly journals Dissecting the function of the different chitin synthases in vegetative growth and sexual development in Neurospora crassa

2015 ◽  
Vol 75 ◽  
pp. 30-45 ◽  
Author(s):  
Rosa A. Fajardo-Somera ◽  
Bastian Jöhnk ◽  
Özgür Bayram ◽  
Oliver Valerius ◽  
Gerhard H. Braus ◽  
...  
Keyword(s):  
Neurospora Crassa ◽  
Sexual Development ◽  
Vegetative Growth ◽  
Chitin Synthases

scholarly journals A Putative Rhamnogalacturonase Required for Sexual Development of Neurospora crassa

Genetics ◽  
1997 ◽  
Vol 146 (2) ◽  
pp. 531-540 ◽  
Author(s):  
Mary Anne Nelson ◽  
Sandra T Merino ◽  
Robert L Metzenberg
Keyword(s):  
Neurospora Crassa ◽  
Filamentous Fungus ◽  
Sexual Development ◽  
Mitotic Division ◽  
Sexual Cycle ◽  
Wild Type ◽  
Aspergillus Aculeatus ◽  
Large Numbers ◽  
Sexual Progeny

In previous work, the asd-1 (ascus development) gene of the filamentous fungus Neurospora crassa was identified as a gene expressed preferentially during the sexual cycle and shown to be essential for normal sexual development. The asd-1 gene has been sequenced and further characterized. It contains two introns, the first of which is in-frame and inefficiently or differentially spliced. The predicted ASD-1 protein has extensive homology with rhamnogalacturonase B of Aspergillus aculeatus, which cleaves the backbone within the ramified hairy regions of pectin. In homozygous asd-1 crosses, sexual development is initiated and large numbers of normal-sized asci are formed. Ascospore delineation does not occur, however, and no sexual progeny are produced. As most asd-1 asci contain eight nuclei, the two meiotic divisions and subsequent mitotic division typical of normal crosses seem to occur, but the haploid nuclei are not partitioned into ascospores. In wild-type crosses, the ASD-1 protein is present in large amounts in croziers and young asci, but it is only faintly detectable in more mature asci containing developing ascospores. Models to explain the possible role of a rhamnogalacturonase in sexual development are presented.

scholarly journals Molecular Characterization of tol, a Mediator of Mating-Type-Associated Vegetative Incompatibility in Neurospora crassa

Genetics ◽  
1999 ◽  
Vol 151 (2) ◽  
pp. 545-555 ◽  
Author(s):  
Patrick Ka Tai Shiu ◽  
N Louise Glass
Keyword(s):  
Sexual Reproduction ◽  
Neurospora Crassa ◽  
Mating Type ◽  
Vegetative Growth ◽  
Point Mutations ◽  
Coiled Coil ◽  
Wild Type ◽  
Opposite Mating Type ◽  
Type Locus

Abstract The mating-type locus in the haploid filamentous fungus, Neurospora crassa, controls mating and sexual development. The fusion of reproductive structures of opposite mating type, A and a, is required to initiate sexual reproduction. However, the fusion of hyphae of opposite mating type during vegetative growth results in growth inhibition and cell death, a process that is mediated by the tol locus. Mutations in tol are recessive and suppress mating-type-associated heterokaryon incompatibility. In this study, we describe the cloning and characterization of tol. The tol gene encodes a putative 1011-amino-acid polypeptide with a coiled-coil domain and a leucine-rich repeat. Both regions are required for tol activity. Repeat-induced point mutations in tol result in mutants that are wild type during vegetative growth and sexual reproduction, but that allow opposite mating-type individuals to form a vigorous heterokaryon. Transcript analyses show that tol mRNA is present during vegetative growth but absent during a cross. These data suggest that tol transcription is repressed to allow the coexistence of opposite mating-type nuclei during the sexual reproductive phase. tol is expressed in a mat A, mat a, A/a partial diploid and in a mating-type deletion strain, indicating that MAT A-1 and MAT a-1 are not absolutely required for transcription or repression of tol. These data suggest that TOL may rather interact with MAT A-1 and/or MAT a-1 (or downstream products) to form a death-triggering complex.

Ultrastructure and Morphology of the Neurospora Crassa Cell Wall Mutants, Slime And Slime X, Using Tem and Sem

1976 ◽  
Vol 34 ◽  
pp. 54-55
Author(s):  
Karen S. Howard ◽  
H. D. Braymer ◽  
M. D. Socolofsky ◽  
S. A. Milligan
Keyword(s):  
Cell Wall ◽  
Neurospora Crassa ◽  
Wild Type ◽  
Morphological Comparison ◽  
Small Areas ◽  
Solid Media ◽  
Thin Sectioning ◽  
X Cells ◽  
Mutant Cells ◽  
Isolated Cell

The recently isolated cell wall mutant slime X of Neurospora crassa was prepared for ultrastructural and morphological comparison with the cell wall mutant slime. The purpose of this article is to discuss the methods of preparation for TEM and SEM observations, as well as to make a preliminary comparison of the two mutants.TEM: Cells of the slime mutant were prepared for thin sectioning by the method of Bigger, et al. Slime X cells were prepared in the same manner with the following two exceptions: the cells were embedded in 3% agar prior to fixation and the buffered solutions contained 5% sucrose throughout the procedure.SEM: Two methods were used to prepare mutant and wild type Neurospora for the SEM. First, single colonies of mutant cells and small areas of wild type hyphae were cut from solid media and fixed with OSO4 vapors similar to the procedure used by Harris, et al. with one alteration. The cell-containing agar blocks were dehydrated by immersion in 2,2-dimethoxypropane (DMP).

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