Antifungal activity of proteolytic fraction (P1G10) from (Vasconcellea cundinamarcensis) latex inhibit cell growth and cell wall integrity in Botrytis cinerea

Abstract Wsc1p, Wsc2p, Wsc3p, and Wsc4p, members of a novel protein family in the yeast Saccharomyces cerevisiae, are putative sensors or receptors in the stress response. Genetic characterization suggests that the WSC family are upstream regulators of the stress-activated PKC1-MAP kinase cascade and are required for the heat shock response and for maintenance of cell wall integrity. The Wsc proteins share sequence characteristics: at their N terminus they have a cysteine motif and a serine/threonine-rich domain predicted to be extracellular, a hydrophobic domain suggested to be transmembranous, and a variable, highly charged C terminus that may be involved in intracellular signaling. Although a role for the WSC genes in maintenance of cell wall integrity has been firmly established, little is known about the properties of the proteins. As reported here, to study its properties in vivo, we epitope tagged the Wsc1 protein. Wsc1p was found to fractionate with the membrane pellet after high-speed centrifugation. Extraction experiments show that Wsc1p is an integral membrane protein present in two forms: one solubilized by detergent, the other Triton X-100 insoluble. Our results also show that Wsc1p is glycosylated and phosphorylated. To characterize the contribution of different domains to the function of Wsc1p, we generated various deletion constructs. Analysis of the properties and function of the mutant proteins shows that the predicted extracellular serine/threonine-rich domain is required for Wsc1p functionality, as well as its glycosylation. A mutant Wsc1 protein lacking the putative transmembrane domain is not functional and partitions to the soluble fraction. Overexpression of full-length Wsc1p inhibits cell growth, with the N terminus alone being sufficient for this inhibition. This suggests that Wsc1p may function in a complex with at least one other protein important for normal cell growth.

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Antifungal Activity against Botrytis cinerea of 2,6-Dimethoxy-4-(phenylimino)cyclohexa-2,5-dienone Derivatives

Molecules ◽

10.3390/molecules24040706 ◽

2019 ◽

Vol 24 (4) ◽

pp. 706 ◽

Cited By ~ 3

Author(s):

Paulo Castro ◽

Leonora Mendoza ◽

Claudio Vásquez ◽

Paz Pereira ◽

Freddy Navarro ◽

...

Keyword(s):

Mass Spectrometry ◽

Cell Wall ◽

Antifungal Activity ◽

Botrytis Cinerea ◽

Aromatic Ring ◽

13C Nmr ◽

Trametes Versicolor ◽

Fungal Cell Wall ◽

Fungal Cell ◽

1H And 13C Nmr

In this work the enzyme laccase from Trametes versicolor was used to synthetize 2,6-dimethoxy-4-(phenylimino)cyclohexa-2,5-dienone derivatives. Ten products with different substitutions in the aromatic ring were synthetized and characterized using 1H- and 13C-NMR and mass spectrometry. The 3,5-dichlorinated compound showed highest antifungal activity against the phytopathogen Botrytis cinerea, while the p-methoxylated compound had the lowest activity; however, the antifungal activity of the products was higher than the activity of the substrates of the reactions. Finally, the results suggested that these compounds produced damage in the fungal cell wall.

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LRR-extensins of vegetative tissues are a functionally conserved family of RALF1 receptors interacting with the receptor kinase FERONIA

10.1101/783266 ◽

2019 ◽

Author(s):

Aline Herger ◽

Shibu Gupta ◽

Gabor Kadler ◽

Christina Maria Franck ◽

Aurélien Boisson-Dernier ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Cell Wall ◽

Plasma Membrane ◽

Cell Growth ◽

Mechanical Stability ◽

Root Hairs ◽

Cell Wall Integrity ◽

Receptor Kinase ◽

Growth Processes ◽

Evolutionary Diversification

AbstractPlant cell growth requires the coordinated expansion of the protoplast and the cell wall that confers mechanical stability to the cell. An elaborate system of cell wall integrity sensors monitors cell wall structures and conveys information on cell wall composition and growth factors to the cell. LRR-extensins (LRXs) are cell wall-attached extracellular regulators of cell wall formation and high-affinity binding sites for RALF (rapid alkalinization factor) peptide hormones that trigger diverse physiological processes related to cell growth. RALF peptides are also perceived by receptors at the plasma membrane and LRX4 of Arabidopsis thaliana has been shown to also interact with one of these receptors, FERONIA (FER). Here, we demonstrate that several LRXs, including the main LRX protein of root hairs, LRX1, interact with FER and RALF1 to coordinate growth processes. Membrane association of LRXs correlate with binding to FER, indicating that LRXs represent a physical link between intra- and extracellular compartments via interaction with membrane-localized proteins. Finally, despite evolutionary diversification of the LRR domains of various LRX proteins, many of them are functionally still overlapping, indicative of LRX proteins being central players in regulatory processes that are conserved in very different cell types.Author SummaryCell growth in plants requires the coordinated enlargement of the cell and the surrounding cell wall, which is ascertained by an elaborate system of cell wall integrity sensors, proteins involved in the exchange of information between the cell and the cell wall. In Arabidopsis thaliana, LRR-extensins (LRXs) are localized in the cell wall and are binding RALF peptides, hormones that regulate cell growth-related processes. LRX4 also binds the plasma membrane-localized receptor kinase FERONIA (FER), establishing a link between the cell and the cell wall. It is not clear, however, whether the different LRXs of Arabidopsis have similar functions and how they interact with their binding partners. Here, we demonstrate that interaction with FER and RALFs requires the LRR domain of LRXs and several but not all LRXs can bind these proteins. This explains the observation that mutations in several of the LRXs induce phenotypes comparable to a fer mutant, establishing that LRX-FER interaction is important for proper cell growth. Some LRXs, however, appear to influence cell growth processes in different ways, which remain to be identified.

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Mkh1, a MEK kinase required for cell wall integrity and proper response to osmotic and temperature stress in Schizosaccharomyces pombe.

Molecular and Cellular Biology ◽

10.1128/mcb.17.7.3508 ◽

1997 ◽

Vol 17 (7) ◽

pp. 3508-3519 ◽

Cited By ~ 50

Author(s):

A S Sengar ◽

N A Markley ◽

N J Marini ◽

D Young

Keyword(s):

Cell Cycle ◽

Cell Wall ◽

Cell Growth ◽

Stationary Phase ◽

Cell Morphology ◽

Cell Shape ◽

Catalytic Domain ◽

Salt Resistance ◽

Cell Wall Integrity ◽

Phase Arrest

We have identified a Schizosaccharomyces pombe gene, mkh1, that encodes a MEK kinase (MEKK) homolog. The coding region of mkh1 is contained within a single exon encoding a 1,116-amino-acid protein. The putative catalytic domain of Mkh1 is 54% identical to the catalytic domain of S. cerevisiae Bck1, the most closely related protein. Deletion of mkh1 did not significantly affect cell growth or division under standard conditions. However, mkh1delta cell growth was inhibited by high KCl or NaCl concentrations. mkh1delta cells required a longer time to reenter the cell cycle after prolonged stationary-phase arrest. Also, mkh1delta cells exhibited a round cell shape, while overexpression of Mkh1 resulted in an elongated cell shape. mkh1delta cells exhibited a more dramatic phenotype when grown in nutrient-limiting conditions at high temperature or in hyperosmotic medium. In such conditions, completion of cytokinesis was inhibited, resulting in the growth of pseudohyphal filaments with multiple septa and nuclei. Also, mkh1delta cells were hypersensitive to beta-glucanase treatment. Together these results suggest that Mkh1 regulates cell morphology, cell wall integrity, salt resistance, cell cycle reentry from stationary-phase arrest, and filamentous growth in response to stress. These phenotypes are essentially identical to those exhibited by cells lacking Pmk1/Spm1, a recently identified mitogen-activated protein kinase. Our evidence suggests that Pmk1/Spm1 acts downstream from Mkh1 in a common pathway. Our results also suggest that Mkh1 and Pck2 act independently to maintain cell wall integrity, cell morphology, and salt resistance but act in opposition to regulate filamentous growth.

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Cinnamaldehyde Exerts Its Antifungal Activity by Disrupting the Cell Wall Integrity of Geotrichum citri-aurantii

Frontiers in Microbiology ◽

10.3389/fmicb.2019.00055 ◽

2019 ◽

Vol 10 ◽

Cited By ~ 18

Author(s):

Qiuli OuYang ◽

Xiaofang Duan ◽

Lu Li ◽

Nengguo Tao

Keyword(s):

Cell Wall ◽

Antifungal Activity ◽

Cell Wall Integrity

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bcpmr1 encodes a P-type Ca2+/Mn2+-ATPase mediating cell-wall integrity and virulence in the phytopathogen Botrytis cinerea

Fungal Genetics and Biology ◽

10.1016/j.fgb.2015.01.012 ◽

2015 ◽

Vol 76 ◽

pp. 36-46 ◽

Cited By ~ 14

Author(s):

Verónica Plaza ◽

Yanssuy Lagües ◽

Mauro Carvajal ◽

Luis A. Pérez-García ◽

Hector M. Mora-Montes ◽

...

Keyword(s):

Cell Wall ◽

Botrytis Cinerea ◽

Cell Wall Integrity ◽

P Type

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Tryptophan recovers sensitivity to cell membrane stress inSaccharomyces cerevisiae

10.1101/344366 ◽

2018 ◽

Author(s):

Lea Schroeder ◽

Pauletta Lazarevskiy ◽

Amy E. Ikui

Keyword(s):

Cell Wall ◽

Cell Growth ◽

Sodium Dodecyl Sulfate ◽

Membrane Damage ◽

Sodium Dodecyl ◽

Cell Wall Integrity ◽

Membrane Stress ◽

Calcofluor White ◽

Functional Link ◽

Dodecyl Sulfate

AbstractSodium dodecyl sulfate is a detergent that disrupts cell membranes, activates cell wall integrity signaling and restricts cell growth inSaccharomyces cerevisiae. However, the underlying mechanism of how sodium dodecyl sulfate inhibits cell growth is not fully understood. Because deletion of theMCK1gene leads to sensitivity to sodium dodecyl sulfate, we implemented a suppressor gene screening revealing that theTAT2tryptophan permease rescues cell growth to sodium dodecyl sulfate-treatedΔmck1cells. Therefore, we questioned the involvement of tryptophan in the response to sodium dodecyl sulfate treatment. In this work, we show thatΔtrp1cells have a disadvantage in the response to sodium dodecyl sulfate compared to auxotrophy for adenine, histidine, leucine or uracil. While also critical in the response to tea tree oil,TRP1does not avert growth inhibition due to other cell wall/membrane perturbations that activate cell wall integrity signaling such as calcofluor white, Congo Red or heat stress. This implicates a distinction from the cell wall integrity pathway and suggests specificity to membrane stress as opposed to cell wall stress. We discover that tyrosine biosynthesis is also essential upon sodium dodecyl sulfate perturbation whereas phenylalanine biosynthesis appears dispensable. Finally, we observe enhanced tryptophan import within minutes upon exposure to sodium dodecyl sulfate indicating that these cells are not starved for tryptophan. In summary, our results expose a functional link between internal tryptophan levels and tryptophan biosynthesis in the response to plasma membrane damage.

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