Komagataella phaffii YPS1-5 encodes the alpha-factor degrading protease Bar1

2020 ◽  
Vol 20 (3) ◽  
Author(s):  
Lina Heistinger ◽  
Brigitte Gasser ◽  
Diethard Mattanovich
Keyword(s):  
Mating Type ◽  
Methylotrophic Yeast ◽  
Sexual Cycle ◽  
Protease Gene ◽  
Komagataella Phaffii ◽  
Highly Sensitive ◽  
Surface Receptor ◽  
Haploid Species ◽  
Secreted Peptides ◽  
Mating Conditions

ABSTRACT Yeast mating pheromones are small secreted peptides required for efficient mating between cells of opposite mating type. Pheromone gradients allow the cells to detect potential mating partners. Secreted pheromone degrading proteases steepen local gradients and allow fast recovery from the pheromone signal. The methylotrophic yeast Komagataella phaffii is a preferentially haploid species. Only under nitrogen starvation, mating genes are activated and the cells are able to undergo a full sexual cycle of mating and sporulation. It has been shown that, similar to other yeasts, K. phaffii requires the mating pheromone and pheromone surface receptor genes for efficient mating. The analysis of so far uncharacterized mating-type-specific genes allowed us to identify the K. phaffii α-factor protease gene YPS1–5. It encodes an aspartic protease of the yapsin family and is upregulated only in a-type cells under mating conditions. The phenotype of K. phaffiia-type strains with a deletion in the protease gene was found to be highly similar to the phenotype of Saccharomyces cerevisiae α-factor protease BAR1 deletion strains. They are highly sensitive to α-factor pheromone in pheromone sensitivity assays and were found to mate with reduced efficiency. Based on our results, we propose to rename the gene into K. phaffii BAR1.

scholarly journals THE SEARCH FOR MATING-TYPE-LIMITED GENES IN THE HOMOTHALLIC ALGA CHLAMYDOMONAS MONOICA

Genetics ◽  
1986 ◽  
Vol 113 (3) ◽  
pp. 601-619
Author(s):  
Karen P VanWinkle-Swift ◽  
Jang-Hee Hahn
Keyword(s):  
Mating Type ◽  
Resistance Mutation ◽  
Mendelian Inheritance ◽  
Sexual Cycle ◽  
Uniparental Inheritance ◽  
Cell Type ◽  
Erythromycin Resistance ◽  
Type Locus ◽  
Heterothallic Species ◽  
Chlamydomonas Monoica

ABSTRACT The non-Mendelian erythromycin resistance mutation ery-u1 shows bidirectional uniparental inheritance in crosses between homothallic ery-u1 and ery-u1  + strains of Chlamydomonas monoica. This inheritance pattern supports a general model for homothallism invoking intrastrain differentiation into opposite compatible mating types and, further, suggests that non-Mendelian inheritance is under mating-type (mt) control in C. monoica as in heterothallic species. However, the identification of genes expressed or required by one gametic cell type, but not the other, is essential to verify the existence of a regulatory mating-type locus in C. monoica and to understand its role in cell differentiation and sexual development. By screening for a shift from bidirectional to unidirectional transmission of the non-Mendelian ery-u1 marker, a mutant with an apparent mating-type-limited sexual cycle defect was obtained. The responsible mutation, mtl-1, causes a 1000-fold reduction in zygospore germination in populations homozygous for the mutant allele and, approximately, a 50% reduction in germination for heterozygous (mtl-1/mtl-1  +) zygospores. By next screening for strains unable to yield any viable zygospores in a cross to mtl-1, a second putative mating-type-limited mutant, mtl-2, was obtained. The mtl-2 strain, although self-sterile, mates efficiently with mtl-2  + strains and shows a unidirectional uniparental pattern of inheritance for the ery-u1 cytoplasmic marker, similar to that observed for crosses involving mtl-1. Genetic analysis indicates that mtl-1 and mtl-2 define unique unlinked Mendelian loci and that the sexual cycle defects of reduced germination (mtl-1) or self-sterility (mtl-2) cosegregate with the effect on ery-u1 cytoplasmic gene transmission. By analogy to C. reinhardtii, the mtl-1 and mtl-2 phenotypes can be explained if the expression of these gene loci is limited to the mt  + gametic cell type, or if the wild-type alleles at these loci are required for the normal formation and/or functioning of mt  + gametes only.

scholarly journals Isolation of Neurospora crassa A mating type mutants by repeat induced point (RIP) mutation.

Genetics ◽  
1992 ◽  
Vol 132 (1) ◽  
pp. 125-133 ◽  
Author(s):  
N L Glass ◽  
L Lee
Keyword(s):  
Neurospora Crassa ◽  
Mating Type ◽  
Single Copy ◽  
Open Reading Frame ◽  
Sexual Cycle ◽  
Reading Frame ◽  
Heterokaryon Incompatibility ◽  
Functional Regions ◽  
A Genome ◽  
Ascospore Formation

Abstract In the filamentous fungus, Neurospora crassa, mating type is regulated by a single locus with alternate alleles, termed A and a. The mating type alleles control entry into the sexual cycle, but during vegetative growth they function to elicit heterokaryon incompatibility, such that fusion of A and a hypha results in death of cells along the fusion point. Previous studies have shown that the A allele consists of 5301 bp and has no similarity to the a allele; it is found as a single copy and only within the A genome. The a allele is 3235 bp in length and it, too, is found as a single copy within the a genome. Within the A sequence, a single open reading frame (ORF) of 288 amino acids (mt A-1) is thought to confer fertility and heterokaryon incompatibility. In this study, we have used repeat induced point (RIP) mutation to identify functional regions of the A idiomorph. RIP mutations in mt A-1 resulted in the isolation of sterile, heterokaryon-compatible mutants, while RIP mutations generated in a region outside of mt A-1 resulted in the isolation of mutants capable of mating, but deficient in ascospore formation.

scholarly journals Population Genetic Analysis of Phytophthora parasitica From Tobacco in Chongqing, Southwestern China

Plant Disease ◽  
2019 ◽  
Vol 103 (10) ◽  
pp. 2599-2605 ◽  
Author(s):  
Qiang Zhang ◽  
Ruirui Feng ◽  
Qing Zheng ◽  
Jinyang Li ◽  
Zhirou Liu ◽  
...  
Keyword(s):  
Genetic Analysis ◽  
Mating Type ◽  
Population Genetic ◽  
Population Analysis ◽  
Genotypic Diversity ◽  
Control Measures ◽  
Economic Losses ◽  
Phytophthora Parasitica ◽  
Population Genetic Analysis ◽  
Highly Sensitive

Tobacco black shank, caused by Phytophthora parasitica, is one of the most notorious tobacco diseases and causes huge economic losses worldwide. Understanding the genetic variation of P. parasitica populations is essential to the development of disease control measures. In this research, 210 simple sequence repeat (SSR) markers for P. parasitica were identified, 10 of which were polymorphic among nine reference strains. We further performed population genetic analysis of 245 P. parasitica isolates randomly collected from tobacco fields in Chongqing for mating type, molecular variation at 14 SSR loci (four of which were identified previously), and sensitivity to the fungicide metalaxyl. The results showed that the A2 mating type was dominant and no A1 mating type isolate was discovered. SSR genotyping distinguished 245 P. parasitica isolates into 46 genotypes, four of which were dominant in the population. Low genotypic diversity and excess heterozygosity were common in nearly all of the populations from Chongqing. Population analysis showed that no differentiation existed among different populations. All isolates tested were highly sensitive to metalaxyl. Taken together, our results showed that the P. parasitica populations from tobacco fields in Chongqing belonged to a clonal lineage and were highly sensitive to metalaxyl.

Fructose‐1,6‐bisphosphatase degradation in the methylotrophic yeast Komagataella phaffii occurs in autophagy pathway

2020 ◽  
Author(s):  
Olena Dmytruk ◽  
Nina Bulbotka ◽  
Anastasya Zazulya ◽  
Marta Semkiv ◽  
Kostyantyn Dmytruk ◽  
...  
Keyword(s):  
Methylotrophic Yeast ◽  
Komagataella Phaffii ◽  
Fructose 1,6 Bisphosphatase ◽  
Autophagy Pathway

scholarly journals Diversity of Phytophthora capsici in Northwest Spain: Analysis of Virulence, Metalaxyl Response, and Molecular Characterization

Plant Disease ◽  
2006 ◽  
Vol 90 (9) ◽  
pp. 1135-1142 ◽  
Author(s):  
C. Silvar ◽  
F. Merino ◽  
J. Díaz
Keyword(s):  
Mating Type ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Phytophthora Capsici ◽  
Crown Rot ◽  
Group Method ◽  
Pair Group ◽  
Highly Sensitive ◽  
Virulence Assay ◽  
Pepper Cultivar

Phytophthora crown rot, caused by Phytophthora capsici, is potentially the most destructive disease of pepper in Spain. Phenotypic and genetic diversity of 16 P. capsici isolates collected from 11 farms in northwest Spain was characterized based on virulence, mating type, sensitivity to metalaxyl, and genetic analysis using random amplified polymorphic DNA (RAPD) methods. Low variability was observed among the isolates in their metalaxyl response, with 87.5% being highly sensitive. No isolates of the A2 mating type were detected. More variability was found in the virulence assay, and isolates were classified into two groups according to their pathogenicity on a set of four pepper cultivar differentials. Genetic variation examined with eight RAPD primers generated 92 polymorphic bands and revealed the existence of different patterns among isolates. Cluster analysis using the unweighted pair-group method with arithmetic averages (UPGMA) separated the Spanish isolates into three RAPD groups and established a relationship between the Spanish population and a representative worldwide group of isolates. No correlation was found between groups obtained by RAPD analysis and groups defined by virulence or metalaxyl response.

Identifying plant cell-surface receptors: combining ‘classical’ techniques with novel methods

2014 ◽  
Vol 42 (2) ◽  
pp. 395-400 ◽  
Author(s):  
Susanne Uebler ◽  
Thomas Dresselhaus
Keyword(s):  
Cell Surface ◽  
Affinity Purification ◽  
Cell Communication ◽  
Membrane Receptors ◽  
Cell Surface Receptor ◽  
Target Cells ◽  
Surface Receptors ◽  
Success Stories ◽  
Surface Receptor ◽  
Secreted Peptides

Cell–cell communication during development and reproduction in plants depends largely on a few phytohormones and many diverse classes of polymorphic secreted peptides. The peptide ligands are bound at the cell surface of target cells by their membranous interaction partners representing, in most cases, either receptor-like kinases or ion channels. Although knowledge of both the extracellular ligand and its corresponding receptor(s) is necessary to describe the downstream signalling pathway(s), to date only a few ligand–receptor pairs have been identified. Several methods, such as affinity purification and yeast two-hybrid screens, have been used very successfully to elucidate interactions between soluble proteins, but most of these methods cannot be applied to membranous proteins. Experimental obstacles such as low concentration and poor solubility of membrane receptors, as well as instable transient interactions, often hamper the use of these ‘classical’ approaches. However, over the last few years, a lot of progress has been made to overcome these problems by combining classical techniques with new methodologies. In the present article, we review the most promising recent methods in identifying cell-surface receptor interactions, with an emphasis on success stories outside the field of plant research.

scholarly journals Identification of the Mating-Type (MAT) Locus That Controls Sexual Reproduction of Blastomyces dermatitidis

2012 ◽  
Vol 12 (1) ◽  
pp. 109-117 ◽  
Author(s):  
Wenjun Li ◽  
Thomas D. Sullivan ◽  
Eric Walton ◽  
Anna Floyd Averette ◽  
Sharadha Sakthikumar ◽  
...  
Keyword(s):  
Sexual Reproduction ◽  
Mating Type ◽  
Genetic Recombination ◽  
Sexual Cycle ◽  
Blastomyces Dermatitidis ◽  
Comparative Genomic ◽  
Dimorphic Fungi ◽  
Content Type ◽  
Long Time ◽  
Northern United States

ABSTRACTBlastomyces dermatitidisis a dimorphic fungal pathogen that primarily causes blastomycosis in the midwestern and northern United States and Canada. While the genes controlling sexual development have been known for a long time, the genes controlling sexual reproduction ofB. dermatitidis(teleomorph,Ajellomyces dermatitidis) are unknown. We identified the mating-type (MAT) locus in theB. dermatitidisgenome by comparative genomic approaches. TheB. dermatitidis MATlocus resembles those of other dimorphic fungi, containing either an alpha-box (MAT1-1) or an HMG domain (MAT1-2) gene linked to theAPN2,SLA2, andCOX13genes. However, in some strains ofB. dermatitidis, theMATlocus harbors transposable elements (TEs) that make it unusually large compared to theMATlocus of other dimorphic fungi. Based on theMATlocus sequences ofB. dermatitidis, we designed specific primers for PCR determination of the mating type. TwoB. dermatitidisisolates of opposite mating types were cocultured on mating medium. Immature sexual structures were observed starting at 3 weeks of coculture, with coiled-hyphae-containing cleistothecia developing over the next 3 to 6 weeks. Genetic recombination was detected in potential progeny by mating-type determination, PCR-restriction fragment length polymorphism (PCR-RFLP), and random amplification of polymorphic DNA (RAPD) analyses, suggesting that a meiotic sexual cycle might have been completed. The F1 progeny were sexually fertile when tested with strains of the opposite mating type. Our studies provide a model for the evolution of theMATlocus in the dimorphic and closely related fungi and open the door to classic genetic analysis and studies on the possible roles of mating and mating type in infection and virulence.

scholarly journals Description of Komagataella phaffii sp. nov. and the transfer of Pichia pseudopastoris to the methylotrophic yeast genus Komagataella

2005 ◽  
Vol 55 (2) ◽  
pp. 973-976 ◽  
Author(s):  
Cletus P. Kurtzman
Keyword(s):  
Type Strain ◽  
Yeast Species ◽  
Large Subunit ◽  
Methylotrophic Yeast ◽  
Ascomycetous Yeast ◽  
26S Rdna ◽  
Komagataella Phaffii ◽  
Oak Trees ◽  
Unidentified Source ◽  
Black Oak

The new methanol-assimilating yeast species Komagataella phaffii Kurtzman sp. nov. (type strain NRRL Y-7556T=CBS 2612T) is described. Of the four known strains of this species, two were isolated from black oak trees in California, USA, one from an Emory oak in Arizona, USA, and one from an unidentified source in Mexico. The species forms hat-shaped ascospores in deliquescent asci and appears to be homothallic. Analysis of nucleotide sequences from domains D1/D2 of large-subunit (26S) rDNA separates the new species from Komagataella pastoris, the type species of the genus, and from Pichia pseudopastoris, which is here renamed Komagataella pseudopastoris (Dlauchy, Tornai-Lehoczki, Fülöp & Péter) Kurtzman comb. nov. (type strain NRRL Y-27603T=CBS 9187T=NCAIM Y 01541T). On the basis of D1/D2 26S rDNA sequence analysis, the three species now assigned to the genus Komagataella represent a clade that is phylogenetically isolated from other ascomycetous yeast genera.

scholarly journals First description of the sexual stage ofVenturia effusa, causal agent of pecan scab

2019 ◽  
Author(s):  
Nikki D. Charlton ◽  
Mihwa Yi ◽  
Clive H. Bock ◽  
Minling Zhang ◽  
Carolyn A. Young
Keyword(s):  
Mating Type ◽  
Culture Media ◽  
Spatial Scales ◽  
Sexual Cycle ◽  
Economic Losses ◽  
Mating Types ◽  
Sexual Stage ◽  
Pecan Scab ◽  
Prevalent Disease ◽  
The Impact

ABSTRACTVenturia effusa, cause of pecan scab, is the most prevalent disease of pecan in the southeastern USA; epidemics of the disease regularly result in economic losses to the pecan industry. Recent characterization of the mating type distribution revealed the frequency of theMATidiomorphs are in equilibrium at various spatial scales, indicative of regular sexual recombination. However, the occurrence of the sexual stage ofV. effusahas never been observed, and the pathogen was previously believed to rely entirely on asexual reproduction. To explore the existence of a sexual cycle, we paired opposite mating types on oatmeal culture media. In initial experiments, cultures were incubated at 24 C for 2 mo for hyphal interactions to occur between mating types and then maintained at 4 C for 4 mo. Immature pseudothecia were initially observed but following exposure to a 12 h photoperiod for 2 weeks at 24 C, asci and ascospores developed. Further experiments explored the effect of time on pseudothecial development with 4 mo at 4 C as the optimal requirement. The results of this study demonstrate the heterothallic nature ofV. effusa.Following experiments investigated progeny from a sexual cross of an albino and a wild-type isolate. Evaluation of isolate pigmentation, mating type, and multilocus genotyping of single ascospore progeny provided evidence that recombination occurred within the sexual crosses. The impact of determining the source of the overwintering ascostroma will aid in management decisions to reduce the primary inoculum in the disease cycle.

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