Identification and characterization of the Komagataella phaffii mating pheromone genes

Abstract The methylotrophic yeast Komagataella phaffii (Pichia pastoris) is a haploid yeast that is able to form diploid cells by mating once nitrogen becomes limiting. Activation of the mating response requires the secretion of a- and α-factor pheromones, which bind to G-protein coupled receptors on cells of opposite mating type. In K. phaffii, the genes coding for the α-factor (MFα), the pheromone surface receptors and the conserved a-factor biogenesis pathway have been annotated previously. Initial homology-based search failed to identify potential a-factor genes (MFA). By using transcriptome data of heterothallic strains under mating conditions, we found two K. phaffiia-factor genes. Deletion of both MFA genes prevented mating of a-type cells. MFA single mutants were still able to mate and activate the mating response pathway in α-type cells. A reporter assay was used to confirm the biological activity of synthetic a- and α-factor peptides. The identification of the a-factor genes enabled the first characterization of the role and regulation of the mating pheromone genes and the response of K. phaffii to synthetic pheromones and will help to gain a better understanding of the mating behavior of K. phaffii.

• Publication year

  2018

• Venue

  FEMS Yeast Research

• Publication date

  2018-04-26

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1093/femsyr/foy051PMID 29718186PMCID 5993090
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• A reporter assay was used to confirm the biological activity of synthetic a- and alpha-factor peptides.

  Confidence 0.98

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review
• Komagataella phaffii MFA single mutants were still able to mate and activate the mating response pathway in alpha-type cells.

  Confidence 0.97

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review
• Deletion of both Komagataella phaffii MFA genes prevented mating of a-type cells.

  Confidence 0.99

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review
• Transcriptome data from heterothallic strains under mating conditions led to the identification of two Komagataella phaffii MFA genes.

  Confidence 0.98

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review

• a-factor pheromone

  pheromone

  The mating pheromone secreted by a-type cells in K. phaffii.

  Aliases: a-factor

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review
• alpha-factor pheromone

  pheromone

  The mating pheromone secreted by alpha-type cells in K. phaffii.

  Aliases: alpha-factor, α-factor

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review
• komagataella phaffii

  organism, yeast

  A methylotrophic yeast species also known as Pichia pastoris.

  Aliases: K. phaffii, Pichia pastoris

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review
• mating response pathway

  pathway

  The signaling pathway that mediates pheromone-driven mating responses.

  Aliases: mating response

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review
• mfa genes

  gene family

  Genes in K. phaffii that encode the a-factor pheromone.

  Aliases: a-factor genes, MFA

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review
• reporter assay

  assay

  An assay that uses a reporter readout to measure biological activity.

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review
• synthetic a- and alpha-factor peptides

  peptide

  Chemically synthesized versions of the a- and alpha-factor pheromones prepared for testing.

  Aliases: synthetic a- and α-factor peptides

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review
• transcriptome data

  dataset

  RNA expression data collected from heterothallic strains under mating conditions and used to search for MFA genes.

  Aliases: transcriptome

  뀨 (7c402c1b98) extractionAnonymous (12632b8b5f) review

• GoldenPiCS: a Golden Gate-derived modular cloning system for applied synthetic biology in the yeast Pichia pastoris

  2017cited by this paper
• Creation of Stable Heterothallic Strains of Komagataella phaffii Enables Dissection of Mating Gene Regulation

  2017cited by this paper
• An efficient tool for metabolic pathway construction and gene integration for Aspergillus niger.

  2017cited by this paper
• Near-optimal probabilistic RNA-seq quantification

  2016influential reference
• Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown

  2016cited by this paper
• Comparative genomics and transcriptomics of Pichia pastoris

  2016cited by this paper
• Curation of the genome annotation of Pichia pastoris (Komagataella phaffii) CBS7435 from gene level to protein function.

  2016cited by this paper
• Molecular coevolution of a sex pheromone and its receptor triggers reproductive isolation in Schizosaccharomyces pombe

  2015cited by this paper
• Orchestrating high-throughput genomic analysis with Bioconductor

  2015cited by this paper
• Near-optimal RNA-Seq quantification

  2015cited by this paper
• Differential analyses for RNA-seq: transcript-level estimates improve gene-level inferences

  2015cited by this paper
• RNA-Seq workflow: gene-level exploratory analysis and differential expression

  2015cited by this paper
• Mating-type switching by chromosomal inversion in methylotrophic yeasts suggests an origin for the three-locus Saccharomyces cerevisiae system

  2014cited by this paper
• R: A language and environment for statistical computing.

  2014cited by this paper
• TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions

  2013influential reference
• Genetically Engineered Transvestites Reveal Novel Mating Genes in Budding Yeast

  2013cited by this paper
• Mate and fuse: how yeast cells do it

  2013cited by this paper
• Pichia pastoris: protein production host and model organism for biomedical research.

  2013cited by this paper
• An improved short‐lived fluorescent protein transcriptional reporter for Saccharomyces cerevisiae

  2012cited by this paper
• Facile synthesis of budding yeast a‐factor and its use to synchronize cells of α mating type

  2012cited by this paper
• Biogenesis of the Saccharomyces cerevisiae Pheromone a-Factor, from Yeast Mating to Human Disease

  2012cited by this paper
• Analysis of changes in protein level and subcellular localization during cell cycle progression using the budding yeast Saccharomyces cerevisiae.

  2011cited by this paper
• Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega

  2011cited by this paper
• Fungal mating pheromones: choreographing the dating game.

  2011cited by this paper
• Characterization of α-factor pheromone and pheromone receptor genes of Ashbya gossypii.

  2011cited by this paper
• A Modular Cloning System for Standardized Assembly of Multigene Constructs

  2011cited by this paper
• High-quality genome sequence of Pichia pastoris CBS7435.

  2011influential reference
• Systematic discovery of unannotated genes in 11 yeast species using a database of orthologous genomic segments

  2011cited by this paper
• Identification and characterisation of novel Pichia pastoris promoters for heterologous protein production.

  2010cited by this paper
• JBrowse: a next-generation genome browser.

  2009cited by this paper
• Kluyveromyces lactis sexual pheromones. Gene structures and cellular responses to alpha-factor.

  2007cited by this paper
• Kluyveromyces lactis sexual pheromones. Gene structures and cellular responses to α-factor

  2007cited by this paper
• Classical genetics.

  2007cited by this paper
• Identification and Characterization of MFA1, the Gene Encoding Candida albicansa-Factor Pheromone

  2007influential reference
• A CHARACTERIZATION OF FIBRANT SEGAL CATEGORIES

  2006cited by this paper
• Unique Aspects of Gene Expression during Candida albicans Mating and Possible G1 Dependency

  2005cited by this paper
• What was classical genetics?

  2004cited by this paper
• Bioconductor: open software development for computational biology and bioinformatics

  2004cited by this paper
• Gene Expression Omnibus: NCBI gene expression and hybridization array data repository

  2002cited by this paper
• Alpha-factor synchronization of budding yeast.

  1997cited by this paper
• New vectors for combinatorial deletions in yeast chromosomes and for gap‐repair cloning using ‘split‐marker’ recombination

  1996cited by this paper
• Analysis of the structural genes encoding M-factor in the fission yeast Schizosaccharomyces pombe: identification of a third gene, mfm3

  1994cited by this paper
• The fission yeast mating pheromone P-factor: its molecular structure, gene structure, and ability to induce gene expression and G1 arrest in the mating partner.

  1994cited by this paper
• Molecular monitoring of the transcriptional activation of the yeast Saccharomyces kluyveri mating pheromone signal transduction by using FUS1-lacZ fusion gene.

  1992cited by this paper
• Mating pheromones of the fission yeast Schizosaccharomyces pombe: purification and structural characterization of M‐factor and isolation and analysis of two genes encoding the pheromone.

  1992cited by this paper
• Significance of C-terminal cysteine modifications to the biological activity of the Saccharomyces cerevisiae a-factor mating pheromone

  1991cited by this paper
• Adenylyl cyclase is dispensable for vegetative cell growth in the fission yeast Schizosaccharomyces pombe.

  1990cited by this paper
• Oligotrophic properties of heterotrophic bacteria and in situ heterotrophic activity in pelagic seawates

  1990cited by this paper
• Saccharomyces cerevisiae STE6 gene product: a novel pathway for protein export in eukaryotic cells.

  1989cited by this paper
• Total synthesis of the lipopeptide a-mating factor of Saccharomyces cerevisiae.

  1989cited by this paper
• Regulation of alpha-factor production in Saccharomyces cerevisiae: a-factor pheromone-induced expression of the MF alpha 1 and STE13 genes

  1989cited by this paper
• The a-factor pheromone of Saccharomyces cerevisiae is essential for mating

  1988influential reference
• The Saccharomyces cerevisiae BAR1 gene encodes an exported protein with homology to pepsin.

  1988cited by this paper
• Structure of Saccharomyces cerevisiae mating hormone a-factor. Identification of S-farnesyl cysteine as a structural component.

  1988cited by this paper
• Two genes required for cell fusion during yeast conjugation: evidence for a pheromone-induced surface protein

  1987cited by this paper
• Identification and regulation of a gene required for cell fusion during mating of the yeast Saccharomyces cerevisiae

  1987cited by this paper
• Amino acid sequences of a-factor mating peptides from Saccharomyces cerevisiae.

  1987cited by this paper
• Induction of yeast mating pheromone a-factor by α cells

  1983cited by this paper
• Saccharomyces cerevisiae contains two discrete genes coding for the alpha-factor pheromone.

  1983cited by this paper
• Induction of yeast mating pheromone a-factor by alpha cells.

  1983cited by this paper
• Structure of a yeast pheromone gene (MF alpha): a putative alpha-factor precursor contains four tandem copies of mature alpha-factor.

  1982cited by this paper
• Structure of a yeast pheromone gene (MFα): A putative α-factor precursor contains four tandem copies of mature α-factor

  1982cited by this paper
• Control of yeast cell type by the mating type locus. I. Identification and control of expression of the a-specific gene BAR1.

  1981cited by this paper
• a-Factor from Saccharomyces cerevisiae: partial characterization of a mating hormone produced by cells of mating type a

  1977cited by this paper

• Decoding the Genome of the Komagataella phaffii Type Strain CBS 2612

  2025influential citation
• The Construction of Heterothallic Strains of Komagataella kurtzmanii Using the I-SceI Meganuclease

  2025cites this paper
• Knock-out of the major regulator Flo8 in Komagataella phaffii results in unique host strain performance for methanol-free recombinant protein production.

  2024cites this paper
• Advances in Metabolic Engineering of Pichia pastoris Strains as Powerful Cell Factories

  2023cites this paper
• Extending the reach of homology by using successive computational filters to find yeast pheromone genes.

  2023cites this paper
• Structural and functional investigation of ABC transporter STE6-2p from Pichia pastoris reveals unexpected interaction with sterol molecules

  2022cites this paper
• Beyond the reach of homology: successive computational filters find yeast pheromone genes

  2021cites this paper
• Komagataella phaffii as Emerging Model Organism in Fundamental Research

  2021cites this paper
• Fermenting Futures: an artistic view on yeast biotechnology.

  2021cites this paper
• Hungry for Sex: Differential Roles for Ustilago maydis b Locus Components in Haploid Cells vis à vis Nutritional Availability

  2021cites this paper
• Komagataella phaffii YPS1-5 encodes the alpha-factor degrading protease Bar1.

  2020cites this paper
• Tetrad analysis without tetrad dissection: Meiotic recombination and genomic diversity in the yeast Komagataella phaffii (Pichia pastoris)

  2019cites this paper
• Selecting for Altered Substrate Specificity Reveals the Evolutionary Flexibilityof ATP-Binding Cassette Transporters

  2019cites this paper
• Genomic diversity and meiotic recombination among isolates of the biotech yeast Komagataella phaffii (Pichia pastoris)

  2019cites this paper
• Carbon source requirements for mating and mating‐type switching in the methylotrophic yeasts Ogataea (Hansenula) polymorpha and Komagataella phaffii (Pichia pastoris)

  2019cites this paper
• Pichia pastoris protease‐deficient and auxotrophic strains generated by a novel, user‐friendly vector toolbox for gene deletion

  2019cites this paper