Cell Wall Trapping of Autocrine Peptides for Human G-Protein-Coupled Receptors on the Yeast Cell Surface

G protein-coupled receptors (GPCRs) are cell surface proteins which play a key role in allowing cells, tissues, and organs to respond to changes in the external environment in order to maintain homeostasis. Despite the fact that GPCRs are known to play key roles in a variety of tissues, there are a large subset of GPCRs that remain poorly studied. In this minireview, we will summarize what is known regarding the “understudied” GPCRs with respect to renal function, and in so doing will highlight the promise represented by studying this gene family.

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Aspergillus fumigatus G-Protein Coupled Receptors GprM and GprJ Are Important for the Regulation of the Cell Wall Integrity Pathway, Secondary Metabolite Production, and Virulence

mBio ◽

10.1128/mbio.02458-20 ◽

2020 ◽

Vol 11 (5) ◽

Cited By ~ 1

Author(s):

Aílton Pereira da Costa Filho ◽

Guilherme Thomaz Pereira Brancini ◽

Patrícia Alves de Castro ◽

Clara Valero ◽

Jaire Alves Ferreira Filho ◽

...

Keyword(s):

Cell Wall ◽

G Protein ◽

Galleria Mellonella ◽

G Protein Coupled Receptors ◽

Cell Wall Integrity ◽

Fungal Development ◽

Content Type ◽

Cell Wall Integrity Pathway ◽

Mycotoxin Biosynthesis ◽

G Protein Coupled

ABSTRACT G-protein coupled receptors (GPCRs) are extracellular signaling receptors that sense environmental cues. Fungi sense their environment primarily through GPCR-mediated signaling pathways, which, in turn, regulate fungal development, metabolism, virulence, and mycotoxin biosynthesis. Aspergillus fumigatus is an important human pathogen that causes aspergillosis, a heterogeneous group of diseases that present a wide range of clinical manifestations. Here, we investigate in detail the role of the GPCRs GprM and GprJ in growth and gene expression. GprM and GprJ are important for melanin production and the regulation of the cell wall integrity (CWI) pathway. Overexpression of gprM and gprJ causes a 20 and 50% reduction in growth rate compared to the wild-type (WT) strain and increases sensitivity to cell wall-damaging agents. Phosphorylation of the CWI protein kinase MpkA is increased in the ΔgprM and ΔgprJ strains and decreased in the overexpression mutants compared to the WT strain. Furthermore, differences in cell wall polysaccharide concentrations and organization were observed in these strains. Transcriptome sequencing suggests that GprM and GprJ negatively regulate genes encoding secondary metabolites (SMs). Mass spectrometry analysis confirmed that the production of fumagillin, pyripyropene, fumigaclavine C, fumiquinazoline, and fumitremorgin is reduced in the ΔgprM and ΔgprJ strains, at least partially through the activation of MpkA. Overexpression of grpM also resulted in the regulation of many transcription factors, with AsgA predicted to function downstream of GprM and MpkA signaling. Finally, we show that the ΔgprM and ΔgprJ mutants are reduced in virulence in the Galleria mellonella insect model of invasive aspergillosis. IMPORTANCE A. fumigatus is the main etiological agent of invasive pulmonary aspergillosis, a life-threatening fungal disease that occurs in severely immunocompromised humans. Withstanding the host environment is essential for A. fumigatus virulence, and sensing of extracellular cues occurs primarily through G-protein coupled receptors (GPCRs) that activate signal transduction pathways, which, in turn, regulate fungal development, metabolism, virulence, and mycotoxin biosynthesis. The A. fumigatus genome encodes 15 putative classical GPCRs, with only three having been functionally characterized to date. In this work, we show that the two GPCRs GprM and GprJ regulate the phosphorylation of the mitogen-activated protein kinase MpkA and thus control the regulation of the cell wall integrity pathway. GprM and GprJ are also involved in the regulation of the production of the secondary metabolites fumagillin, pyripyropene, fumigaclavine C, fumiquinazoline, melanin, and fumitremorgin, and this regulation partially occurs through the activation of MpkA. Furthermore, GprM and GprJ are important for virulence in the insect model Galleria mellonella. This work therefore functionally characterizes two GPCRs and shows how they regulate several intracellular pathways that have been shown to be crucial for A. fumigatus virulence.

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ADP-Ribosylation Factors Modulate the Cell Surface Transport of G Protein-Coupled Receptors

Journal of Pharmacology and Experimental Therapeutics ◽

10.1124/jpet.109.161489 ◽

2010 ◽

Vol 333 (1) ◽

pp. 174-183 ◽

Cited By ~ 32

Author(s):

Chunmin Dong ◽

Xiaoping Zhang ◽

Fuguo Zhou ◽

Huijuan Dou ◽

Matthew T. Duvernay ◽

...

Keyword(s):

Cell Surface ◽

G Protein ◽

G Protein Coupled Receptors ◽

Surface Transport ◽

Adp Ribosylation ◽

G Protein Coupled

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The Endoplasmic Reticulum Ca2+-pump SERCA2b Interacts with G Protein-coupled Receptors and Enhances their Expression at the Cell Surface

Journal of Molecular Biology ◽

10.1016/j.jmb.2007.02.108 ◽

2007 ◽

Vol 371 (3) ◽

pp. 622-638 ◽

Cited By ~ 15

Author(s):

Jussi T. Tuusa ◽

Piia M.H. Markkanen ◽

Pirjo M. Apaja ◽

Anna E. Hakalahti ◽

Ulla E. Petäjä-Repo

Keyword(s):

Endoplasmic Reticulum ◽

Cell Surface ◽

G Protein ◽

G Protein Coupled Receptors ◽

G Protein Coupled

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Oligomerisation of G-protein-coupled receptors

Journal of Cell Science ◽

10.1242/jcs.114.7.1265 ◽

2001 ◽

Vol 114 (7) ◽

pp. 1265-1271 ◽

Cited By ~ 1

Author(s):

G. Milligan

Keyword(s):

Energy Transfer ◽

Cell Surface ◽

G Protein ◽

Living Cells ◽

G Protein Coupled Receptors ◽

Intact Cells ◽

Effective Delivery ◽

Multiple Copies ◽

Agonist Ligands ◽

G Protein Coupled

A range of approaches have recently provided evidence that G-protein-coupled receptors can exist as oligomeric complexes. Both homo-oligomers, comprising multiple copies of the same gene product, and hetero-oligomers containing more than one receptor have been detected. In several, but not all, examples, the extent of oligomerisation is regulated by the presence of agonist ligands, and emerging evidence indicates that receptor hetero-oligomers can display distinct pharmacological characteristics. A chaperonin-like role for receptor oligomerisation in effective delivery of newly synthesised receptors to the cell surface is a developing concept, and recent studies have employed a series of energy-transfer techniques to explore the presence and regulation of receptor oligomerisation in living cells. However, the majority of studies have relied largely on co-immunoprecipitation techniques, and there is still little direct information on the fraction of receptors existing as oligomers in intact cells.

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Cell-Surface Receptors Transactivation Mediated by G Protein-Coupled Receptors

International Journal of Molecular Sciences ◽

10.3390/ijms151119700 ◽

2014 ◽

Vol 15 (11) ◽

pp. 19700-19728 ◽

Cited By ~ 83

Author(s):

Fabio Cattaneo ◽

Germano Guerra ◽

Melania Parisi ◽

Marta De Marinis ◽

Domenico Tafuri ◽

...

Keyword(s):

Cell Surface ◽

G Protein ◽

G Protein Coupled Receptors ◽

Cell Surface Receptors ◽

Surface Receptors ◽

G Protein Coupled

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Structural instability and divergence from conserved residues underlie intracellular retention of mammalian odorant receptors

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1915520117 ◽

2020 ◽

Vol 117 (6) ◽

pp. 2957-2967

Author(s):

Kentaro Ikegami ◽

Claire A. de March ◽

Maira H. Nagai ◽

Soumadwip Ghosh ◽

Matthew Do ◽

...

Keyword(s):

Cell Surface ◽

G Protein ◽

Surface Expression ◽

Structural Instability ◽

G Protein Coupled Receptors ◽

Machine Learning Techniques ◽

Odorant Receptors ◽

Cell Surface Expression ◽

Conserved Residues ◽

G Protein Coupled

Mammalian odorant receptors are a diverse and rapidly evolving set of G protein-coupled receptors expressed in olfactory cilia membranes. Most odorant receptors show little to no cell surface expression in nonolfactory cells due to endoplasmic reticulum retention, which has slowed down biochemical studies. Here we provide evidence that structural instability and divergence from conserved residues of individual odorant receptors underlie intracellular retention using a combination of large-scale screening of odorant receptors cell surface expression in heterologous cells, point mutations, structural modeling, and machine learning techniques. We demonstrate the importance of conserved residues by synthesizing consensus odorant receptors that show high levels of cell surface expression similar to conventional G protein-coupled receptors. Furthermore, we associate in silico structural instability with poor cell surface expression using molecular dynamics simulations. We propose an enhanced evolutionary capacitance of olfactory sensory neurons that enable the functional expression of odorant receptors with cryptic mutations.

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PST1 and ECM33 encode two yeast cell surface GPI proteins important for cell wall integrity

Microbiology ◽

10.1099/mic.0.26924-0 ◽

2004 ◽

Vol 150 (12) ◽

pp. 4157-4170 ◽

Cited By ~ 66

Author(s):

M. Pardo

Keyword(s):

Cell Wall ◽

Cell Surface ◽

Yeast Cell ◽

Cell Wall Integrity ◽

Yeast Cell Surface ◽

Gpi Proteins

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Cell-surface G-protein-coupled receptors for tumor-associated metabolites: A direct link to mitochondrial dysfunction in cancer

Biochimica et Biophysica Acta (BBA) - Reviews on Cancer ◽

10.1016/j.bbcan.2017.05.003 ◽

2017 ◽

Vol 1868 (1) ◽

pp. 246-257 ◽

Cited By ~ 15

Author(s):

Bojana Ristic ◽

Yangzom D. Bhutia ◽

Vadivel Ganapathy

Keyword(s):

Cell Surface ◽

Mitochondrial Dysfunction ◽

G Protein ◽

G Protein Coupled Receptors ◽

Direct Link ◽

G Protein Coupled

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Aspergillus fumigatus G-protein coupled receptors GprM and GprJ are important for the regulation of the cell wall integrity pathway, secondary metabolite production, and virulence

10.1101/2020.08.03.235119 ◽

2020 ◽

Author(s):

Aílton Pereira da Costa Filho ◽

Guilherme Thomaz Pereira Brancini ◽

Patrícia Alves de Castro ◽

Jaire Alves Ferreira ◽

Lilian Pereira Silva ◽

...

Keyword(s):

Cell Wall ◽

Protein Kinase ◽

G Protein ◽

Galleria Mellonella ◽

G Protein Coupled Receptors ◽

Cell Wall Integrity ◽

Fungal Development ◽

Cell Wall Integrity Pathway ◽

Mycotoxin Biosynthesis ◽

G Protein Coupled

AbstractG-protein coupled receptors (GPCRs) are extracellular signalling receptors that sense environmental cues to coordinate a biological response. Fungi sense their environment primarily through GPCR-mediated signalling pathways, which in turn regulate fungal development, metabolism, virulence and mycotoxin biosynthesis. A. fumigatus is an important human pathogen that causes aspergillosis, a heterogeneous group of diseases that presents a wide range of clinical manifestations. Here, we investigate in detail the role of the GPCRs GprM and GprJ in growth and gene expression. GprM and GprJ are important for melanin production and the regulation of the cell wall integrity (CWI) pathway. Overexpression of gprM and gprJ causes a 20 and 50% reduction in growth rate when compared to the wild-type (WT) strain, and increases sensitivity to cell wall-damaging agents. Phosphorylation of the CWI protein kinase MpkA is increased in the ΔgprM and ΔgprJ strains and decreased in the overexpression mutants when compared to the WT strain. Furthermore, differences in cell wall polysaccharide concentrations and organization were observed in these strains. RNA-sequencing suggests that GprM and GprJ negatively regulate genes encoding secondary metabolites (SMs). Mass spectrometry analysis confirmed that the production of fumagillin, pyripyropene, fumigaclavine C, fumiquinazoline, and fumitremorgin is reduced in the ΔgprM and ΔgprJ strains, and that this regulation occurs, at least partially, through the activation of MpkA. Overexpression of grpM also resulted in the regulation of many transcription factors, with AsgA predicted to function downstream of GprM and MpkA signalling. Finally, we show that the ΔgprM and ΔgprJ mutants are reduced in virulence in the Galleria mellonella insect model of invasive aspergillosis. This work further contributes to unravelling functions of A. fumigatus GPCRs and shows that GprM and GprJ are essential for CWI, secondary metabolite production and virulence.Author summaryA. fumigatus is the main ethiological agent of invasive pulmonary aspergillosis, a life-threatening fungal disease that occurs in severely immuno-compromised humans. Withstanding the host environment is essential for A. fumigatus virulence and sensing of extracellular cues occurs primarily through G-protein coupled receptors (GPCRs) that activate signal transduction pathways, which in turn regulate fungal development, metabolism, virulence and mycotoxin biosynthesis. The A. fumigatus genome encodes 15 putative classical GPCRs, with only three having been functionally characterized to date. In this work, we show that the two GPCRs GprM and GprJ regulate the phosphorylation of the mitogen-activated protein kinase MpkA and thus control the regulation of the cell wall integrity pathway. GprM and GprJ are also involved in the regulation of the production of the secondary metabolites fumagillin, pyripyropene, fumigaclavine C, fumiquinazoline, melanin, and fumitremorgin and this regulation partially occurs through the activation of MpkA. Furthermore, GprM and GprJ are important for virulence in the insect model Galleria mellonella. This work therefore functionally characterizes two GPCRs and shows how they regulate several intracellular pathways that have been shown to be crucial for A. fumigatus virulence.

Download Full-text