Importance of MAP Kinases during Protoperithecial Morphogenesis in Neurospora crassa

Lichius, Alexander; Lord, Kathryn M.; Jeffree, C. E.; Oborny, Radek; Boonyarungsrit, Patid; Read, Nick D.

doi:10.1371/journal.pone.0042565

Cited by 40 publications

(55 citation statements)

References 100 publications

(142 reference statements)

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“…1(A)). As previously reported, Δmak-1 showed a rosette-like colony growth, 19,31,40) whereas Δmak-2 exhibited conidiation that started from the colony center. 27,28) These colony phenotypes of Δmak-1 or Δmak-2 were conserved within each MAPK pathway (data not shown), which was also consistent with previous findings.…”

Section: Sensitivity Of Individual Mapk Gene-knockout Strains To Cellsupporting

confidence: 82%

“…27,28) These colony phenotypes of Δmak-1 or Δmak-2 were conserved within each MAPK pathway (data not shown), which was also consistent with previous findings. 19,27,28,31,40) The relative radial growth rates of these disruptants were reduced to less than 40% of that of the wild-type strain grown on agarsolidified VN medium for 24 h at 28°C (Fig. 1(B)).…”

Section: Sensitivity Of Individual Mapk Gene-knockout Strains To Cellmentioning

confidence: 96%

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Involvement of MAK-1 and MAK-2 MAP kinases in cell wall integrity in Neurospora crassa

Kamei

Yamashita

Takahashi

et al. 2016

Bioscience, Biotechnology, and Biochemistry

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Among three MAPK disruptants of Neurospora crassa, Δmak-1 was sensitive and Δmak-2 was hypersensitive to micafungin, a beta-1,3-glucan synthase inhibitor, than the wild-type or Δos-2 strains. We identified six micafungin-inducible genes that are involved in cell wall integrity (CWI) and found that MAK-1 regulated the transcription of non-anchored cell wall protein gene, ncw-1, and the beta-1,3-endoglucanase gene, bgt-2, whereas MAK-2 controlled the expression of the glycosylhydrolase-like protein gene, gh76-5, and the C4-dicarboxylate transporter gene, tdt-1. Western blotting analysis revealed that, in the wild-type strain, MAK-1 was constitutively phosphorylated from conidial germination to hyphal development. In contrast, the phosphorylation of MAK-2 was growth phase-dependent, and micafungin induced the phosphorylation of unphosphorylated MAK-2. It should be noted that the phosphorylation of MAK-1 was virtually abolished in the Δmak-2 strain, but was significantly induced by micafungin, suggesting functional cross talk between MAK-1 and MAK-2 signalling pathway in CWI.

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Section: Sensitivity Of Individual Mapk Gene-knockout Strains To Cellsupporting

confidence: 82%

Section: Sensitivity Of Individual Mapk Gene-knockout Strains To Cellmentioning

confidence: 96%

Involvement of MAK-1 and MAK-2 MAP kinases in cell wall integrity in Neurospora crassa

Kamei

Yamashita

Takahashi

et al. 2016

Bioscience, Biotechnology, and Biochemistry

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“…All knockout strains used in this study were produced and verified, typically using Southern blot analysis, within the NIH Neurospora Genome Knock-Out Project (41), and PCR analysis was used to verify genotypes by following the recently optimized approach of Lichius et al (42). For functional studies of genes, including het-6, het-14, het-15, mkr-5, mkr-6, qde-1, sad-1, sad-3, sms-2, sms-3, and the stc1-like gene, knockouts of both mat a and mat A were available for crossing and phenotyping across sexual development (see Table 2).…”

Section: Methodsmentioning

confidence: 99%

Global Gene Expression and Focused Knockout Analysis Reveals Genes Associated with Fungal Fruiting Body Development in Neurospora crassa

et al. 2014

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Fungi can serve as highly tractable models for understanding genetic basis of sexual development in multicellular organisms. Applying a reverse-genetic approach to advance such a model, we used random and multitargeted primers to assay gene expression across perithecial development in Neurospora crassa. We found that functionally unclassified proteins accounted for most upregulated genes, whereas downregulated genes were enriched for diverse functions. Moreover, genes associated with developmental traits exhibited stage-specific peaks of expression. Expression increased significantly across sexual development for mating type gene mat a-1 and for mat A-1 specific pheromone precursor ccg-4. In addition, expression of a gene encoding a protein similar to zinc finger, stc1, was highly upregulated early in perithecial development, and a strain with a knockout of this gene exhibited arrest at the same developmental stage. A similar expression pattern was observed for genes in RNA silencing and signaling pathways, and strains with knockouts of these genes were also arrested at stages of perithecial development that paralleled their peak in expression. The observed stage specificity allowed us to correlate expression upregulation and developmental progression and to identify regulators of sexual development. Bayesian networks inferred from our expression data revealed previously known and new putative interactions between RNA silencing genes and pathways. Overall, our analysis provides a finescale transcriptomic landscape and novel inferences regarding the control of the multistage development process of sexual crossing and fruiting body development in N. crassa. S hifts in gene expression over the course of development have been attributed a primary role in the unfolding of the developmental program of animals and plants (1-4). However, the genomic basis of development in a third multicellular clade, the fungi, is arguably very different and the least well understood. Estimated as comprising 1.5 to 7.1 million species, fungi have deep evolutionary origins (5-7) and diverse body plans, ranging from highly reduced unicellular species such as microsporidia and yeasts to notoriously large hyphal mats that produce multicellular fruiting bodies such as mushrooms, which feature specialized cell types (8). To address multicellular fruiting body development from a reverse-genetic approach, model fungi can provide ideal systems, as they are easily manipulated, develop fruiting structures with a few well-characterized tissue types, and have relatively small genome sizes, so numerous fungal genomes have been sequenced.Neurospora crassa is a multicellular ascomycete fungus in the family Sordariomycetes, which has been used as a genetic model organism due to its simple filamentous asexual stage (9, 10), and which exhibits promise for revealing the molecular basis of the more complex sexual development of fungi. N. crassa has 28 morphologically distinct cell types, including 14 that are finely differentiated during the development of...

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“…All strains used and produced in this study are listed in supplementary material Table S1. The homokaryotic status of gene deletion mutants was verified by PCR-genotyping as outlined in detail elsewhere (Lichius et al, 2012a). Transformant strains and expression plasmids generated in this study have been deposited at the FGSC for public use; accession numbers are listed in supplementary material Table S2.…”

Section: Strains Transformation and Transformant Selectionmentioning

confidence: 99%

“…MAPKs of the pheromone response (PR) and CWI pathways, but not the HOG pathway, are required for cell fusion in N. crassa (Pandey et al, 2004;Li et al, 2005;Roca et al, 2005;Maerz et al, 2008;Lichius et al, 2012a). The PR pathway MAPK MAK-2 exhibits oscillatory recruitment to CAT tips as they grow chemotropically towards each other.…”

Section: Introductionmentioning

confidence: 99%

CDC-42 and RAC-1 regulate opposite chemotropisms inNeurospora crassa

Lichius

Goryachev

Fricker

et al. 2014

Journal of Cell Science

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Cell polarization and fusion are crucial developmental processes that occur in response to intracellular and extracellular signals. Asexual spores (conidia) of the mold Neurospora crassa differentiate two types of polarized cell protrusions, germ tubes and conidial anastomosis tubes (CATs), which exhibit negative and positive chemotropism, respectively. We provide the first evidence that shared and separate functions of the Rho-type GTPases CDC-42 and RAC-1 regulate these opposite chemotropisms. We demonstrate that RAC-1 is essential for CAT formation and cell fusion, whereas CDC-42 is necessary and sufficient for normal germ tube development. Cdc42-Rac-interactive-binding (CRIB) reporters were constructed to exclusively label locally activated GTP-bound GTPases. Time course analyses showed that repositioning of these activated GTPase clusters within germ tube and CAT tip apices controls directional growth in the absence of a tip-localized vesicle supply center (Spitzenkö rper). We propose a model in which the local assembly of a plasma-membraneassociated GTPase-PAK-MAPK signaling platform regulates chemoattractant perception and secretion in order to synchronize oscillatory cell-cell communication and directional CAT tip growth.

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Importance of MAP Kinases during Protoperithecial Morphogenesis in Neurospora crassa

Cited by 40 publications

References 100 publications

Involvement of MAK-1 and MAK-2 MAP kinases in cell wall integrity in Neurospora crassa

Involvement of MAK-1 and MAK-2 MAP kinases in cell wall integrity in Neurospora crassa

Global Gene Expression and Focused Knockout Analysis Reveals Genes Associated with Fungal Fruiting Body Development in Neurospora crassa

CDC-42 and RAC-1 regulate opposite chemotropisms inNeurospora crassa

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