scholarly journals The rice blast fungus MoRgs1 functioning in cAMP signaling and pathogenicity is regulated by casein kinase MoCk2 phosphorylation and modulated by membrane protein MoEmc2

2021 ◽  
Vol 17 (6) ◽  
pp. e1009657
Author(s):  
Rui Yu ◽  
Xuetong Shen ◽  
Muxing Liu ◽  
Xinyu Liu ◽  
Ziyi Yin ◽  
...  
Keyword(s):  
Membrane Protein ◽  
G Protein ◽  
Rice Blast ◽  
Casein Kinase ◽  
Camp Signaling ◽  
Normal Operation ◽  
G Protein Signaling ◽  
Protein Signaling ◽  
Regulatory Subunits ◽  
Genes Encoding

GTP-binding protein (G-protein) and regulator of G-protein signaling (RGS) mediated signal transduction are critical in the growth and virulence of the rice blast pathogen Magnaporthe oryzae. We have previously reported that there are eight RGS and RGS-like proteins named MoRgs1 to MoRgs8 playing distinct and shared regulatory functions in M. oryzae and that MoRgs1 has a more prominent role compared to others in the fungus. To further explore the unique regulatory mechanism of MoRgs1, we screened a M. oryzae cDNA library for genes encoding MoRgs1-interacting proteins and identified MoCkb2, one of the two regulatory subunits of the casein kinase (CK) 2 MoCk2. We found that MoCkb2 and the sole catalytic subunit MoCka1 are required for the phosphorylation of MoRgs1 at the plasma membrane (PM) and late endosome (LE). We further found that an endoplasmic reticulum (ER) membrane protein complex (EMC) subunit, MoEmc2, modulates the phosphorylation of MoRgs1 by MoCk2. Interestingly, this phosphorylation is also essential for the GTPase-activating protein (GAP) function of MoRgs1. The balance among MoRgs1, MoCk2, and MoEmc2 ensures normal operation of the G-protein MoMagA-cAMP signaling required for appressorium formation and pathogenicity of the fungus. This has been the first report that an EMC subunit is directly linked to G-protein signaling through modulation of an RGS-casein kinase interaction.

scholarly journals Heterotrimeric G-Protein Signaling Is Required for Cellulose Degradation in Neurospora crassa

mBio ◽  
2020 ◽  
Vol 11 (6) ◽  
Author(s):  
Logan A. Collier ◽  
Arit Ghosh ◽  
Katherine A. Borkovich
Keyword(s):  
G Protein ◽  
Cellulose Degradation ◽  
Cellulase Activity ◽  
Cellulase Production ◽  
Camp Signaling ◽  
G Protein Signaling ◽  
Protein Signaling ◽  
Wild Type ◽  
Posttranscriptional Control ◽  
Heterotrimeric G Protein Signaling

ABSTRACT The filamentous fungus Neurospora crassa decomposes lignocellulosic biomass to generate soluble sugars as carbon sources. In this study, we investigated a role for heterotrimeric G-protein signaling in cellulose degradation. Loss of the Gα subunit genes gna-1 and gna-3, the Gβ subunit genes gnb-1 and cpc-2, the Gγ gene gng-1, or the gene for downstream effector adenylyl cyclase (cr-1) resulted in loss of detectable cellulase activity. This defect was also observed in strains expressing a constitutively active version of gna-3 (gna-3Q208L). We found that GNA-1 levels are greatly reduced in Δgna-3, Δgnb-1, and Δgng-1 strains, likely contributing to cellulase defects in these genetic backgrounds. The observation that gna-3Q208L Δgnb-1 strains exhibit cellulase activity, despite greatly reduced levels of GNA-1 protein, is consistent with positive control of cellulase production by GNA-3 that is manifested in the absence of gnb-1. Expression patterns for five cellulase genes showed that Δgna-1, Δgnb-1, and Δgna-3 mutants produce less cellulase mRNA than the wild type, consistent with transcriptional regulation. Δcpc-2 mutants had wild-type levels of cellulase transcripts, suggesting posttranscriptional control. In contrast, results for Δcr-1 mutants support both transcriptional and posttranscriptional control of cellulase activity by cAMP signaling. Cellulase activity defects in Δgna-3 mutants were fully remediated by cAMP supplementation, consistent with GNA-3 operating upstream of cAMP signaling. In contrast, cAMP addition only partially corrected cellulase activity defects in Δgna-1 and Δgnb-1 mutants, suggesting participation of GNA-1 and GNB-1 in additional cAMP-independent pathways that control cellulase activity. IMPORTANCE Filamentous fungi are critical for the recycling of plant litter in the biosphere by degrading lignocellulosic biomass into simpler compounds for metabolism. Both saprophytic and pathogenic fungi utilize plant cell wall-degrading enzymes to liberate carbon for metabolism. Several studies have demonstrated a role for cellulase enzymes during infection of economically relevant crops by fungal pathogens. Especially in developing countries, severe plant disease means loss of entire crops, sometimes leading to starvation. In this study, we demonstrate that G-protein signaling is a key component of cellulase production. Therefore, understanding the role of G-protein signaling in the regulation of the unique metabolism of cellulose by these organisms can inform innovations in strain engineering of industrially relevant species for biofuel production and in combatting food shortages caused by plant pathogens.

scholarly journals The RGS Protein Crg2 Regulates Pheromone and Cyclic AMP Signaling in Cryptococcus neoformans

2008 ◽  
Vol 7 (9) ◽  
pp. 1540-1548 ◽  
Author(s):  
Gui Shen ◽  
Yan-Li Wang ◽  
Amy Whittington ◽  
Lie Li ◽  
Ping Wang
Keyword(s):  
Cyclic Amp ◽  
Cryptococcus Neoformans ◽  
G Protein ◽  
Mitogen Activated Protein Kinase ◽  
Camp Signaling ◽  
Cell Swelling ◽  
G Protein Signaling ◽  
Protein Signaling ◽  
Protein Activity

ABSTRACT Crg1 and Crg2 are regulators of G-protein signaling homologs found in the human fungal pathogen Cryptococcus neoformans. Crg1 negatively regulates pheromone responses and mating through direct inhibition of Gα subunits Gpa2 and Gpa3. It has also been proposed that Crg2 has a role in mating, as genetic crosses involving Δcrg2 mutants resulted in formation of hyperfilaments. We found that mutation of Gpa2 and Gpa3 partially suppressed the hyperfilamentation, mutation of Gpa3 alleviated Δcrg2-specfic cell swelling, and mutation of the mitogen-activated protein kinase Cpk1 blocked both processes. These findings indicate that Gpa2 and Gpa3 function downstream of Crg2 and that Gpa3 is also epistatic to Crg2 in a Cpk1-dependent morphogenesis process linked to mating. Significantly, we found that Δcrg2 mutants formed enlarged capsules that mimic cells expressing a constitutively active GPA1(Q284L) allele and that the levels of intracellular cyclic AMP (cAMP) were also elevated, suggesting that Crg2 also negatively regulates the Gpa1-cAMP signaling pathway. We further showed that Crg2 interacted with Gpa3 and Gpa1, but not Gpa2, in a pulldown assay and that Crg2 maintained a higher in vitro GTPase-activating protein activity toward Gpa3 and Gpa1 than to Gpa2. Finally, we found that dysregulation of cAMP due to the Crg2 mutation attenuated virulence in a murine model of cryptococcosis. Taken together, our study reveals Crg2 as an RGS (regulator of G-protein signaling) protein of multiregulatory function, including one that controls mating distinctly from Crg1 and one that serves as a novel inhibitor of Gpa1-cAMP signaling.

scholarly journals MoRic8 Is a Novel Component of G-Protein Signaling During Plant Infection by the Rice Blast Fungus Magnaporthe oryzae

2010 ◽  
Vol 23 (3) ◽  
pp. 317-331 ◽  
Author(s):  
Ya Li ◽  
Xia Yan ◽  
Hong Wang ◽  
Shen Liang ◽  
Wei-Bin Ma ◽  
...  
Keyword(s):  
G Protein ◽  
Magnaporthe Oryzae ◽  
Rice Blast ◽  
Insertional Mutagenesis ◽  
Rice Blast Fungus ◽  
G Protein Signaling ◽  
Protein Signaling ◽  
Targeted Gene Deletion ◽  
Plant Infection ◽  
Blast Fungus

An insertional mutagenesis screen was used to investigate the biology of plant infection by the devastating rice blast pathogen, Magnaporthe oryzae. Here, we report the identification of a new mutant, LY-130, which is defective in multiple steps during infection-related morphogenesis and pathogenicity. Analysis of the mutation confirmed an insertion into gene MoRIC8, which encodes a 480-amino-acid protein that is a putative homologue of the Ric8 regulator of GTP-binding protein (G-protein) signaling, previously described in animals. Targeted gene deletion mutants of MoRIC8 were nonpathogenic and impaired in cellular differentiation associated with sporulation, sexual development, and plant infection. MoRic8 physically interacts with the Gα subunit MagB in yeast two-hybrid assays and appears to act upstream of the cyclic AMP response pathway that is necessary for appressorium morphogenesis. Taken together, our results indicate that MoRic8 may act as a novel regulator of the G-protein signaling during infection-related development of rice blast fungus M. oryzae.

scholarly journals RgsA Attenuates the PKA Signaling, Stress Response, and Virulence in the Human Opportunistic Pathogen Aspergillus fumigatus

2019 ◽  
Vol 20 (22) ◽  
pp. 5628 ◽  
Author(s):  
Hnin Phyu Lwin ◽  
Yong-Ho Choi ◽  
Min-Woo Lee ◽  
Jae-Hyuk Yu ◽  
Kwang-Soo Shin
Keyword(s):  
Stress Response ◽  
Aspergillus Fumigatus ◽  
G Protein ◽  
Stress Responses ◽  
Pathogenic Fungus ◽  
G Protein Signaling ◽  
Mrna Levels ◽  
Protein Signaling ◽  
Null Mutant ◽  
Genes Encoding

The regulator of G-protein signaling (RGS) proteins play an important role in upstream control of heterotrimeric G-protein signaling pathways. In the genome of the human opportunistic pathogenic fungus Aspergillus fumigatus, six RGS protein-encoding genes are present. To characterize the rgsA gene predicted to encode a protein with an RGS domain, we generated an rgsA null mutant and observed the phenotypes of the mutant. The deletion (Δ) of rgsA resulted in increased radial growth and enhanced asexual sporulation in both solid and liquid culture conditions. Accordingly, transcripts levels of the key asexual developmental regulators abaA, brlA, and wetA are elevated in the ΔrgsA mutant. Moreover, ΔrgsA resulted in elevated spore germination rates in the absence of a carbon source. The activity of cAMP-dependent protein kinase A (PKA) and mRNA levels of genes encoding PKA signaling elements are elevated by ΔrgsA. In addition, mRNA levels of genes associated with stress-response signaling increased with the lack of rgsA, and the ΔrgsA spores showed enhanced tolerance against oxidative stressors. Comparative transcriptomic analyses revealed that the ΔrgsA mutant showed higher mRNA levels of gliotoxin (GT) biosynthetic genes. Accordingly, the rgsA null mutant exhibited increased production of GT and elevated virulence in the mouse. Conversely, the majority of genes encoding glucan degrading enzymes were down-regulated by ΔrgsA, and endoglucanase activities were reduced. In summary, RgsA plays multiple roles, governing growth, development, stress responses, virulence, and external polymer degradation—likely by attenuating PKA signaling.

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