ESCAPE FROM MATING-TYPE INCOMPATIBILITY IN BISEXUAL (A + a) NEUROSPORA HETEROKARYONS

1975 ◽  
Vol 17 (3) ◽  
pp. 441-449 ◽  
Author(s):  
A. M. DeLange ◽  
A. J. F. Griffiths
Keyword(s):  
Mating Type ◽  
Loss Of Function ◽  
Wild Type ◽  
Opposite Mating Type ◽  
Type Locus ◽  
Heterokaryon Incompatibility ◽  
Recessive Mutant ◽  
Type Rate ◽  
Wild Type Rate ◽  
Incompatibility Locus

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.

A SEARCH FOR COMPLEXITY AT THE MATING-TYPE LOCUS OF NEUROSPORA CRASSA

1973 ◽  
Vol 15 (3) ◽  
pp. 577-585 ◽  
Author(s):  
Dorothy Newmeyer ◽  
H. Branch Howe Jr. ◽  
Donna R. Galeazzi
Keyword(s):  
Neurospora Crassa ◽  
Mating Type ◽  
Adjacent Gene ◽  
Opposite Mating Type ◽  
Type Locus ◽  
Linked Gene ◽  
Heterokaryon Incompatibility ◽  
Mating Type Locus ◽  
Incompatibility Reaction ◽  
The Right

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.

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.

scholarly journals Functional Redundancy in the Hydroxycinnamate Catabolism Pathways of the Salt Marsh BacteriumSagittula stellataE-37

2018 ◽  
Vol 84 (23) ◽  
Author(s):  
Ashley M. Frank ◽  
Michelle J. Chua ◽  
Christopher A. Gulvik ◽  
Alison Buchan
Keyword(s):  
Marine Bacterium ◽  
Vascular Plant ◽  
Functional Redundancy ◽  
Bacterial Degradation ◽  
Degradation Pathways ◽  
Wild Type ◽  
Content Type ◽  
Essential Components ◽  
Type Rate ◽  
Wild Type Rate

ABSTRACTThe hydroxycinnamates (HCAs) ferulate andp-coumarate are among the most abundant constituents of lignin, and their degradation by bacteria is an essential step in the remineralization of vascular plant material. Here, we investigate the catabolism of these two HCAs by the marine bacteriumSagittula stellataE-37, a member of the roseobacter lineage with lignolytic potential. Bacterial degradation of HCAs is often initiated by the activity of a hydroxycinnamoyl-coenzyme A (hydroxycinnamoyl-CoA) synthase. Genome analysis ofS. stellatarevealed the presence of two feruloyl-CoA (fcs) synthase homologs, an unusual occurrence among characterized HCA degraders. In order to elucidate the role of these homologs in HCA catabolism,fcs-1andfcs-2were disrupted using insertional mutagenesis, yielding both single and doublefcsmutants. Growth onp-coumarate was abolished in thefcsdouble mutant, whereas maximum cell yield on ferulate was only 2% of that of the wild type. Interestingly, the single mutants demonstrated opposing phenotypes, where thefcs-1mutant showed impaired growth (extended lag and ∼60% of wild-type rate) onp-coumarate, and thefcs-2mutant showed impaired growth (extended lag and ∼20% of wild-type rate) on ferulate, pointing to distinct but overlapping roles of the encodedfcshomologs, withfcs-1primarily dedicated top-coumarate utilization andfcs-2playing a dominant role in ferulate utilization. Finally, a tripartite ATP-independent periplasmic (TRAP) family transporter was found to be required for growth on both HCAs. These findings provide evidence for functional redundancy in the degradation of HCAs inS. stellataE-37 and offer important insight into the genetic complexity of aromatic compound degradation in bacteria.IMPORTANCEHydroxycinnamates (HCAs) are essential components of lignin and are involved in various plant functions, including defense. In nature, microbial degradation of HCAs is influential to global carbon cycling. HCA degradation pathways are also of industrial relevance, as microbial transformation of the HCA, ferulate, can generate vanillin, a valuable flavoring compound. Yet, surprisingly little is known of the genetics underlying bacterial HCA degradation. Here, we make comparisons to previously characterized bacterial HCA degraders and use a genetic approach to characterize genes involved in catabolism and uptake of HCAs in the environmentally relevant marine bacteriumSagittula stellata. We provide evidence of overlapping substrate specificity between HCA degradation pathways and uptake proteins. We conclude thatS. stellatais uniquely poised to utilize HCAs found in the complex mixtures of plant-derived compounds in nature. This strategy may be common among marine bacteria residing in lignin-rich coastal waters and has potential relevance to biotechnology sectors.

scholarly journals LINKAGE OF MUTATIONS AFFECTING MINUS FLAGELLAR MEMBRANE AGGLUTINABILITY TO THE mt  - MATING-TYPE LOCUS OF CHLAMYDOMONAS

Genetics ◽  
1981 ◽  
Vol 99 (1) ◽  
pp. 41-47 ◽  
Author(s):  
Carol J Hwang ◽  
Brian C Monk ◽  
Ursula W Goodenough
Keyword(s):  
Uv Irradiation ◽  
Mating Type ◽  
Zygote Formation ◽  
Wild Type ◽  
Tetrad Analysis ◽  
Type Locus ◽  
Mating Type Locus ◽  
Flagellar Membrane ◽  
Mutant Strains

ABSTRACT Two independently isolated mutant strains, imp-10 and imp-12, were obtained by UV irradiation of wild-type mating-type minus (wt-). Each fails to agglutinate sexually with gametes of either mating type, but mating and zygote formation can be elicited by agglutinating either strain to wt+ gametes by means of anti-flagellar antiserum. Tetrad analysis of the resultant zygotes shows that both imp-10 and imp-12 are very closely linked to mt  -, with no recombinants observed. Diploid strains constructed between imp-10 or imp-12 and wt+ gametes are wt-, that is, they agglutinate and fuse like normal minus cells. Tetrad analysis of triploids from imp-10 diploid x wt+ haploid crosses shows that only imp-10 and wt+ products are recovered. A model is proposed to account for these results.

MUTATIONS OF THE a MATING-TYPE GENE IN NEUROSPORA CRASSA

Genetics ◽  
1978 ◽  
Vol 88 (2) ◽  
pp. 239-254 ◽  
Author(s):  
A J F Griffiths ◽  
A M DeLange
Keyword(s):  
Mating Type ◽  
Mutational Analysis ◽  
Chromosome 1 ◽  
Selection System ◽  
Type Locus ◽  
Heterokaryon Incompatibility ◽  
Mating Type Locus ◽  
Mating Type Gene ◽  
Type Gene ◽  
Restoration Of Fertility

ABSTRACT In Neurospora, the mating-type locus controls both mating (A + a is fertile) and heterokaryosis (A + a is incompatible). The two alleles appear stable: no novel fertility reactions have ever been reported, and attempts to separate fertility and heterokaryon incompatibility functions by recombination have been unsuccessful. In the present approach the locus was studied through a mutational analysis of heterokaryon incompatibility function. A selection system was used that detects vigorous (A + a) heterokaryotic colonies against a background of inhibited growth. Twenty-five mutants of an a strain were produced following mutagenic treatment with UV and NG: 15 were viable as homokaryons and 10 were not. All but one were infertile, but most showed an abortive mating reaction involving the production of barren, well-developed perithecia with A and (surprisingly) a testers. None of the mutants complement each other to restore fertility. Seven mutants have been mapped to the mating-type locus region of chromosome 1. Restoration of fertility was used to detect revertants, and these were found in five out of the eight mutants tested. (A dose response was observed). In four cases incompatibility was fully restored and in one case it was not.—The results suggest two positive actions of the locus when in heterozygous (A/a) combination (the stimulation of some stage of ascus production and the inhibition of vegetative heterokaryosis), and one positive action in homozygous combination (the production of a perithecial inhibitor).

scholarly journals The Pseudomonas aeruginosa devB/SOL Homolog,pgl, Is a Member of the hex Regulon and Encodes 6-Phosphogluconolactonase

2000 ◽  
Vol 182 (14) ◽  
pp. 3934-3941 ◽  
Author(s):  
Paul W. Hager ◽  
M. Worth Calfee ◽  
Paul V. Phibbs
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Carbon Source ◽  
Normal Growth ◽  
Wild Type ◽  
Reading Frame ◽  
Cell Extracts ◽  
Genes Encoding ◽  
Type Rate ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Wild Type Rate

ABSTRACT A cyclic version of the Entner-Doudoroff pathway is used byPseudomonas aeruginosa to metabolize carbohydrates. Genes encoding the enzymes that catabolize intracellular glucose to pyruvate and glyceraldehyde 3-phosphate are coordinately regulated, clustered at 39 min on the chromosome, and collectively form thehex regulon. Within the hex cluster is an open reading frame (ORF) with homology to the devB/SOLfamily of unidentified proteins. This ORF encodes a protein of either 243 or 238 amino acids; it overlaps the 5′ end of zwf (encodes glucose-6-phosphate dehydrogenase) and is followed immediately by eda (encodes the Entner-Doudoroff aldolase). The devB/SOL homolog was inactivated in P. aeruginosa PAO1 by recombination with a suicide plasmid containing an interrupted copy of the gene, creating mutant strain PAO8029. PAO8029 grows at 9% of the wild-type rate using mannitol as the carbon source and at 50% of the wild-type rate using gluconate as the carbon source. Cell extracts of PAO8029 were specifically deficient in 6-phosphogluconolactonase (Pgl) activity. The cloned devB/SOL homolog complemented PAO8029 to restore normal growth on mannitol and gluconate and restored Pgl activity. Hence, we have identified this gene as pgland propose that the devB/SOL family members encode 6-phosphogluconolactonases. Interestingly, three eukaryotic glucose-6-phosphate dehydrogenase (G6PDH) isozymes, from human, rabbit, and Plasmodium falciparum, contain Pgl domains, suggesting that the sequential reactions of G6PDH and Pgl are incorporated in a single protein. 6-Phosphogluconolactonase activity is induced in P. aeruginosa PAO1 by growth on mannitol and repressed by growth on succinate, and it is expressed constitutively in P. aeruginosa PAO8026 (hexR). Taken together, these results establish that Pgl is an essential enzyme of the cyclic Entner-Doudoroff pathway encoded by pgl, a structural gene of the hex regulon.

scholarly journals Effects of Ploidy and Mating Type on Virulence of Candida albicans

2005 ◽  
Vol 73 (11) ◽  
pp. 7366-7374 ◽  
Author(s):  
Ashraf S. Ibrahim ◽  
B. B. Magee ◽  
D. C. Sheppard ◽  
Molly Yang ◽  
Sarah Kauffman ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Mating Type ◽  
Murine Model ◽  
Fungal Pathogen ◽  
Opposite Mating Type ◽  
Type Locus ◽  
Disseminated Candidiasis ◽  
Strain Background ◽  
Do So

ABSTRACT Candida albicans is the most common fungal pathogen of humans. The recent discovery of sexuality in this organism has led to the demonstration of a mating type locus which is usually heterozygous, although some isolates are homozygous. Tetraploids can be formed between homozygotes of the opposite mating type. However, the role of the mating process and tetraploid formation in virulence has not been investigated. We describe here experiments using a murine model of disseminated candidiasis which demonstrate that in three strains, including CAI-4, the most commonly used strain background, tetraploids are less virulent than diploids and can undergo changes in ploidy during infection. In contrast to reports with other strains, we find that MTL homozygotes are almost as virulent as the heterozygotes. These results show that the level of ploidy in Candida albicans can affect virulence, but the mating type configuration does not necessarily do so.

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.

Changes in Mate Recognition Through Alterations of Pheromones and Receptors in the Multisexual Mushroom FungusSchizophyllum commune

Genetics ◽  
2001 ◽  
Vol 158 (4) ◽  
pp. 1491-1503 ◽  
Author(s):  
Thomas J Fowler ◽  
Michael F Mitton ◽  
Lisa J Vaillancourt ◽  
Carlene A Raper
Keyword(s):  
Amino Acid ◽  
Mating Type ◽  
Schizophyllum Commune ◽  
Mate Recognition ◽  
Mating Types ◽  
Loss Of Function ◽  
Sequence Comparisons ◽  
Type Locus ◽  
Step Model ◽  
Genes Encoding

AbstractSchizophyllum commune has thousands of mating types defined in part by numerous lipopeptide pheromones and their G-protein-coupled receptors. These molecules are encoded within multiple versions of two redundantly functioning B mating-type loci, Bα and Bβ. Compatible combinations of pheromones and receptors, produced by individuals of different B mating types, trigger a pathway of fertilization required for sexual development. Analysis of the Bβ2 mating-type locus revealed a large cluster of genes encoding a single pheromone receptor and eight different pheromones. Phenotypic effects of mutations within these genes indicated that small changes in both types of molecules could significantly alter their specificity of interaction. For example, a conservative amino acid substitution in a pheromone resulted in a gain of function toward one receptor and a loss of function with another. A two-amino-acid deletion from a receptor precluded the mutant pheromone from activating the mutant receptor, yet this receptor was activated by other pheromones. Sequence comparisons provided clues toward understanding how so many variants of these multigenic loci could have evolved through duplication and mutational divergence. A three-step model for the origin of new variants comparable to those found in nature is presented.

scholarly journals MUTANTS OF THE KILLER PLASMID OF SACCHAROMYCES CEREVISIAE DEPENDENT ON CHROMOSOMAL DIPLOIDY FOR EXPRESSION AND MAINTENANCE

Genetics ◽  
1976 ◽  
Vol 82 (2) ◽  
pp. 273-285
Author(s):  
Reed B Wickner
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Elevated Temperature ◽  
Mating Type ◽  
Chromosomal Gene ◽  
Wild Type ◽  
Opposite Mating Type ◽  
Diploid Clone ◽  
Wild Type Phenotype ◽  
Chromosomal Genes ◽  
Killer Plasmid

ABSTRACT Mutants of the killer plasmid of Saccharomyecs cerevisiaehave been isolated that depend upon chromosomal diploidy for the expression of plasmid functions and for replication or maintenance of the plasmid itself. These mutants are not defective in any chromosomal gene needed for expression or replication of the killer plasmid.—Haploids carrying these mutant plasmids (called d for diploid-dependent) are either unable to kill or unable to resist being killed or both and show frequent loss of the plasmid. The wild-type phenotype (K+R+) is restored by mating the d plasmid-carrying strain with either (a) a wild-type sensitive strain which apparently has no killer plasmid; (b) a strain which has been cured of the killer plasmid by growth at elevated temperature; (c) a strain which has been cured of the plasmid by growth in the presence of cycloheximide; (d) a strain which has lost the plasmid because it carries a mutation in a chromosomal mak gene; or (e) a strain of the opposite mating type which carries the same d plasmid and has the same defective phenotype, indicating that the restoration of the normal phenotype is not due to recombination between plasmid genomes or complementation of plasmid or chromosomal genes.—Sporulation of the phenotypically K+R+ diploids formed in matings between d and wild-type nonkiller strains yields tetrads, all four of whose haploid spores are defective for killing or resistance or maintenance of the plasmid or a combination of these. Every defective phenotype may be found among the segregants of a single diploid clone carrying a d plasmid. These defective segregants resume the normal killer phenotype in the diploids formed when a second round of mating is performed, and the segregants from a second round of meiosis and sporulation are again defective.

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