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

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.

Genetics ◽  
1998 ◽  
Vol 148 (3) ◽  
pp. 1069-1079 ◽  
Author(s):  
Adlane V-B Ferreira ◽  
Zhiqiang An ◽  
Robert L Metzenberg ◽  
N Louise Glass

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.


1975 ◽  
Vol 17 (3) ◽  
pp. 441-449 ◽  
Author(s):  
A. M. DeLange ◽  
A. J. F. Griffiths

In Neurospora crassa, strains of opposite mating type generally do not form stable heterokaryons because the mating type locus acts as a heterokaryon incompatibility locus. However, when one A and one a strain, having complementing auxotrophic mutants, are placed together on minimal medium, growth may occur, although the growth is generally slow. In this study, escape from such slow growth to that at a wild type or near-wild type rate was observed. The escaped cultures are stable heterokaryons, mostly having lost the mating type allele function from one component nucleus, so that the nuclear types are heterokaryon compatible. Either A or a mating type can be lost. This loss of function has been attributed to deletion since only one nuclear type could be recovered in all heterokaryons except one, but deletion spanning adjacent loci has been directly demonstrated in a minority of cases. Alternatively when one component strain is tol and the other tol+ (tol being a recessive mutant suppressing the heterokaryon incompatibility associated with mating type), escape may occur by the deletion or mutation of tol+, also resulting in heterokaryon compatibility. An induction mechanism for escape is speculated upon.


1973 ◽  
Vol 15 (3) ◽  
pp. 577-585 ◽  
Author(s):  
Dorothy Newmeyer ◽  
H. Branch Howe Jr. ◽  
Donna R. Galeazzi

Evidence for complexity at the mating-type locus of Neurospora crassa was sought by selecting recombinants between closely linked markers on either side. All recombinants were tested for crossing ability, to test the hypothesis that the two mating-type alleles are actually closely linked self-sterile mutants; such tests should also detect subunits analogous to the α and β subunits of the A factor of Schizophyllum or Coprinus. No change in crossing ability was found among the 5,019 recombinants tested, representing 235,000 viable ascospores. The results indicate that if subunits exist, they are not more than 0.002 units apart. Twelve hundred and forty of the recombinants were tested in a way that should also have detected subunits analogous to the A and B factors of Schizophyllum and Coprinus, except that A and B would be closely linked. No such subunits were detected.N. crassa strains of opposite mating type are heterokaryon-incompatible during vegetative growth, and observations of various investigators have suggested that the heterokaryon incompatibility might be controlled by a separate closely-linked gene rather than by mating type itself. A sample of the recombinants was therefore tested for separation of the heterokaryon-incompatibility and crossing-compatibility functions. (Heterokaryon-incompatibility was scored by the presence of an incompatibility reaction in duplications heterozygous for mating type; this technique is simple and eliminates complications due to unlinked heterokaryon-incompatibility loci, several of which are known in N. crassa.) No separation was found. The results indicate that if an adjacent gene is responsible for the heterokaryon-incompatibility, it is not more than 0.0078 units from mating type, if on the left, and not more than 0.018 units from mating type, if on the right.


1970 ◽  
Vol 12 (4) ◽  
pp. 914-926 ◽  
Author(s):  
Dorothy Newmeyer

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.


2019 ◽  
Vol 53 (1) ◽  
pp. 417-444 ◽  
Author(s):  
Sheng Sun ◽  
Marco A. Coelho ◽  
Márcia David-Palma ◽  
Shelby J. Priest ◽  
Joseph Heitman

Cryptococcus species utilize a variety of sexual reproduction mechanisms, which generate genetic diversity, purge deleterious mutations, and contribute to their ability to occupy myriad environmental niches and exhibit a range of pathogenic potential. The bisexual and unisexual cycles of pathogenic Cryptococcus species are stimulated by properties associated with their environmental niches and proceed through well-characterized signaling pathways and corresponding morphological changes. Genes governing mating are encoded by the mating-type ( MAT) loci and influence pathogenesis, population dynamics, and lineage divergence in Cryptococcus. MAT has undergone significant evolutionary changes within the Cryptococcus genus, including transition from the ancestral tetrapolar state in nonpathogenic species to a bipolar mating system in pathogenic species, as well as several internal reconfigurations. Owing to the variety of established sexual reproduction mechanisms and the robust characterization of the evolution of mating and MAT in this genus, Cryptococcus species provide key insights into the evolution of sexual reproduction.


Genetics ◽  
1996 ◽  
Vol 143 (4) ◽  
pp. 1589-1600 ◽  
Author(s):  
Sven J Saupe ◽  
Gretchen A Kuldau ◽  
Myron L Smith ◽  
N Louise Glass

Abstract Filamentous fungi are capable of hyphal fusion, but heterokaryon formation between different isolates is controlled by specific loci termed het loci. Heterokaryotic cells formed between strains of different het genotype are rapidly destroyed or strongly inhibited in their growth. In Neurospora crassa, at least 11 loci, including the mating type locus, affect the capacity to form a heterokaryon between different isolates. In this report, we describe the molecular characterization of the vegetative incompatibility locus, het-C. The het-COR allele was cloned by genetically identifying the het-C locus in a chromosome walk, and the activity of clones containing the het-COR allele was tested in a functional transformation assay. The het-COR allele encodes a 966amino acid polypeptide with a putative signal peptide, a coiled-coil motif and a Gterminal glycine-rich domain, similar to glycine-rich domains detected in various extracellular and structural cell envelope proteins. Both the coiled-coil and one-third of the glycine-rich carboxyl terminal domains were required for full het-COR activity. Mutants of het-COR were obtained by repeat-induced point mutation (RIP) ; these mutants were indistinguishable from wild type during vegetative growth and sexual reproduction hut displayed dual compatibility with both of two mutually incompatible het-COR and het-cPA strains.


2017 ◽  
Vol 49 (4) ◽  
pp. 373-385 ◽  
Author(s):  
Lars R. LUDWIG ◽  
Tina C. SUMMERFIELD ◽  
Janice M. LORD ◽  
Garima SINGH

AbstractIn the present study, we characterized the mating-type locus ofKnightiella splachnirima(Icmadophilaceae) using degenerate and inverse PCR techniques. We screened for the presence of both mating-type locus idiomorphs in DNA extracts of minuscule samples of haploid thalline tissue. We found that only one of the two idiomorphs was present in each sample, and the mating-type ratio (MAT1-1:MAT1-2) was very balanced, being 8:10 and 13:14 at local and global scales, respectively. This indicates that the species is probably self-incompatible and requires the presence of compatible mating partners for sexual reproduction (heterothallic mating system). Furthermore, we provide a mating-type screening protocol withK. splachnirimaspecific mating-type locus primers, which could serve as an essential tool for the conservation management of this rare Australasian endemic.


Genome ◽  
1992 ◽  
Vol 35 (2) ◽  
pp. 347-353 ◽  
Author(s):  
David J. Jacobson

The mating-type of Neurospora crassa (A and a) have a dual function: A and a individuals are required for sexual reproduction, but only strains of the same mating type will form a stable vegetative heterokaryon. Neurospora tetrasperma, in contrast, is a naturally occurring A + a heterokaryon. It was shown previously that the mating-type genes of both species are functionally the same and are not responsible for this difference in heterokaryon incompatibility. This suggests that a separate genetic system determines the heterokaryon incompatibility function of mating type. The mutant tolerant (tol) in N. crassa, unlinked to mating type, acts as a specific suppressor of A + a heterokaryon incompatibility. In the present study, the wild-type alleles at the tol locus were introgressed reciprocally, from N. crassa into N. tetrasperma and from N. tetrasperma into N. crassa, to investigate the action of these alleles in the A + a heterokaryon incompatibility systems of these species. The wild-type allele from N. tetrasperma (tolT) acts as a recessive suppressor of A + a heterokaryon incompatibility in N. crassa. Furthermore, the wild-type allele from N. crassa (tolC) causes A and a to become heterokaryon incompatible in N. tetrasperma, while having no effect on the sexual reproduction. Therefore, the tol gene plays a major role in determining the heterokaryon compatibility of mating type in these species: tolC is an active allele that causes incompatibility and tolT an inactive allele that suppresses incompatibility by its inactivity.Key words: heterokaryon incompatibility, vegetative incompatibility, Neurospora, suppressors, mating type.


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