Off the wall: The rhyme and reason of Neurospora crassa hyphal morphogenesis

Verdín, Jorge; Sánchez‐León, Eddy; Rico-Ramírez, Adriana M.; Martínez‐Núñez, Leonora; Fajardo-Somera, Rosa A.; Riquelme, Meritxell

doi:10.1016/j.tcsw.2019.100020

Cited by 25 publications

(39 citation statements)

References 266 publications

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“…The Neurospora cell wall is comprised of β-Glucans, chitin, galactomannan, and proteins. A compact layer of chitin is situated directly against the plasma membrane (Free, 2013;Verdín et al, 2019). Zymolyase is a common enzymatic preparation used for digestion of cell walls in fungi (Lee et al, 2016;Li and Neuert, 2019).…”

Section: Recombinant Bacillus Licheniformis Chitinase As a Tool For Rnase-free Cell Wall Digestion Of N Crassamentioning

confidence: 99%

Quantitative single molecule RNA-FISH and RNase-free cell wall digestion in Neurospora crassa

Bartholomai

Gladfelter

Loros

et al. 2021

Preprint

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Single molecule RNA-FISH (smFISH) is a valuable tool for analysis of mRNA spatial patterning in fixed cells that is underutilized in filamentous fungi. A primary complication for fixed-cell imaging in filamentous fungi is the need for enzymatic cell wall permeabilization, which is compounded by considerable variability in cell wall composition between species. smFISH adds another layer of complexity due to a requirement for RNase free conditions. Here, we describe the cloning, expression, and purification of a chitinase suitable for supplementation of a commercially available RNase-free enzyme preparation for efficient permeabilization of the Neurospora cell wall. We further provide a method for smFISH in Neurospora which includes a tool for generating numerical data from images that can be used in downstream customized analysis protocols.

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Section: Recombinant Bacillus Licheniformis Chitinase As a Tool For Rnase-free Cell Wall Digestion Of N Crassamentioning

confidence: 99%

Quantitative single molecule RNA-FISH and RNase-free cell wall digestion in Neurospora crassa

Bartholomai

Gladfelter

Loros

et al. 2021

Preprint

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“…Several reviews have summarised the various aspects of polarised growth and hyphal morphogenesis in FF over the past decade (Peñalva 2010 ; Harris 2011 ; Shaw et al 2011 ; Steinberg 2014 ; Takeshita 2016 ; Steinberg et al 2017 ; Riquelme et al 2018 ; Verdín et al 2019 ). For a more comprehensive review on the process of endocytosis itself, see (Steinberg 2014 ; Steinberg et al 2017 ).…”

Section: Overviewmentioning

confidence: 99%

Current views on endocytosis in filamentous fungi

Commer

Shaw

2020

Mycology

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Filamentous fungi grow by adding cell wall and membrane exclusively at the apex of tubular structures called hyphae. Growth was previously believed to occur only through exocytosis at the Spitzenkörper, an organised body of secretory macro-and microvesicles found only in growing hyphae. More recent work has indicated that an area deemed the sub-apical collar is enriched for endocytosis and is also required for hyphal growth. It is now generally believed that polarity of filamentous fungi is achieved through the balancing of the processes of endocytosis and exocytosis at these two areas. This review is an update on the current progress and understanding surrounding the occurrence of endocytosis and its spatial regulation as they pertain to growth and pathogenicity in filamentous fungi.

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“…The morphological state of filamentous fungi also has a large impact on the production of lignocellulases, particularly in submerged or solid‐state fermentations (Cairns et al, 2019; Veiter et al, 2018). A major determinant of cellular morphology is the cell wall, which is regulated by the cell wall integrity (CWI) signaling pathway involving the cell surface sensors such as WSC‐1 and MAP‐kinase‐mediated pathways (Verdín et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Parallel proteomic and phosphoproteomic analyses reveal cellobiose‐dependent regulation of lignocellulase secretion in the filamentous fungus Neurospora crassa

et al. 2021

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High cost of lignocellulases restricts the commercialization of biofuel and bio‐product production from lignocellulosic biomass. Constitutively expressed lignocellulases are considered to degrade cellulose to release small amount of soluble cellodextrins such as cellobiose for further large‐scale production of lignocellulases; however, the underlying mechanism remains to be elucidated. Here, a triple β‐glucosidase mutant of the model fungus Neurospora crassa, which prevents rapid turnover of cellobiose and thus allows the disaccharide to induce lignocellulases, was applied to perform parallel analyses of proteome and phosphoproteome changes in response to cellobiose and Avicel cellulose. The results revealed shared proteome and phosphoproteome responses to cellobiose and Avicel, corroborating the idea that cellobiose mediates the regulation of lignocellulase expression and secretion. The results further suggest that this regulation is achieved at multiple levels, including epigenetic, transcription, post‐transcription, translation, and post‐translation. Proteome profiling revealed that the proteins upregulated by cellobiose and Avicel were over‐represented in cellulose degradation and degradation product transport pathways. Phosphoproteome profiling revealed that the proteins differentially phosphorylated by cellobiose and Avicel were over‐represented by the pathways such as transcriptional control, protein processing and export, cell wall biogenesis, and cellular signaling. Deletion mutation analysis further suggests that the ER chaperon protein Hsp70‐6, the translocation complex subunit Sec66/Sec71, and the signal peptidase subunit Spc2 are involved in lignocellulase secretion, particularly translocation across the endoplasmic reticulum. Altogether, the results offer a new insight into how cellobiose mediates the regulation of lignocellulase expression and secretion, providing a potential strategy for the strain engineering to improve lignocellulase production.

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Off the wall: The rhyme and reason of Neurospora crassa hyphal morphogenesis

Cited by 25 publications

References 266 publications

Quantitative single molecule RNA-FISH and RNase-free cell wall digestion in Neurospora crassa

Quantitative single molecule RNA-FISH and RNase-free cell wall digestion in Neurospora crassa

Current views on endocytosis in filamentous fungi

Parallel proteomic and phosphoproteomic analyses reveal cellobiose‐dependent regulation of lignocellulase secretion in the filamentous fungus Neurospora crassa

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