A secretory phospholipase A2 of a fungal pathogen contributes to lipid droplet homeostasis, assimilation of insect‐derived lipids, and repression of host immune responses

Secretory phospholipase A2s (sPLA2s) are found in a wide range of organisms from bacteria to higher plants and animals and are involved in varied and cellular processes. However, roles of these enzymes in microbial pathogens remain unclear. Here, an sPLA2 (BbPLA2) was characterized in the filamentous insect pathogenic fungus, Beauveria bassiana. BbPLA2 was exclusively expressed in insect hemolymph‐derived cells (hyphal bodies), and its expression was induced by insect‐derived nutrients and lipids, and nutrient starvation. High levels of secretion of BbPLA2 were observed as well as its distribution in hyphal body lipid drops (LDs). Overexpression of BbPLA2 increased the ability of B. bassiana to utilize insect‐derived nutrients and lipids, and promoted LD accumulation, indicating functions for BbPLA2 in mediating LD homeostasis and assimilation of insect‐derived lipids. Strains overexpressing BbPLA2 showed moderately increased virulence, including more efficient penetration of the insect cuticle and evasion of host immune responses as compared to the wild type strain. In addition, B. bassiana‐activated host immune genes were downregulated in the BbPLA2 overexpression strain, but upregulated by infections with a ΔBbPLA2 strain. These data demonstrate that BbPLA2 contributes to LD homeostasis, assimilation of insect‐derived lipids, and repression of host immune responses.

[1]  C. An,et al.  PLA2 mediates the innate immune response in Asian corn borer, Ostrinia furnacalis , 2021, Insect science.

[2]  T. Butt,et al.  RNAi-mediated suppression of insect metalloprotease inhibitor (IMPI) enhances Galleria mellonella susceptibility to fungal infection. , 2021, Developmental and comparative immunology.

[3]  Yongjun Zhang,et al.  Multifunctional role of a fungal pathogen-secreted laccase 2 in evasion of insect immune defense. , 2021, Environmental microbiology.

[4]  A. Meola,et al.  PLA2G1B is involved in CD4 anergy and CD4 lymphopenia in HIV-infected patients. , 2020, The Journal of clinical investigation.

[5]  M. Mehrabadi,et al.  The innate immune gene Relish and Caudal jointly contribute to the gut immune homeostasis by regulating antimicrobial peptides in Galleria mellonella. , 2020, Developmental and comparative immunology.

[6]  Keping Chen,et al.  Beauveria bassiana ribotoxin inhibits insect immunity responses to facilitate infection via host translational blockage. , 2020, Developmental and comparative immunology.

[7]  Yongjun Zhang,et al.  MADS-box transcription factor Mcm1 controls cell cycle, fungal development, cell integrity and virulence in the filamentous insect pathogenic fungus Beauveria bassiana. , 2019, Environmental microbiology.

[8]  Shabbir Ahmed,et al.  A prophylactic role of a secretory PLA2 of Spodoptera exigua against entomopathogens , 2019, Developmental and comparative immunology.

[9]  U. Bora,et al.  Phospholipases play multiple cellular roles including growth, stress tolerance, sexual development, and virulence in fungi. , 2018, Microbiological research.

[10]  Varnavas D. Mouchlis,et al.  Review of four major distinct types of human phospholipase A2. , 2017, Advances in biological regulation.

[11]  N. Keyhani Lipid biology in fungal stress and virulence: Entomopathogenic fungi. , 2017, Fungal biology.

[12]  L. Collinson,et al.  Epithelial-Cell-Derived Phospholipase A2 Group 1B Is an Endogenous Anthelmintic , 2017, Cell Host and Microbe.

[13]  P. Dash,et al.  Molecular details of secretory phospholipase A2 from flax (Linum usitatissimum L.) provide insight into its structure and function , 2017, Scientific Reports.

[14]  H. Han,et al.  Identification and characterization of Vibrio vulnificus plpA encoding a phospholipase A2 essential for pathogenesis , 2017, The Journal of Biological Chemistry.

[15]  Yongjun Zhang,et al.  Correlation of cell surface proteins of distinct Beauveria bassiana cell types and adaption to varied environment and interaction with the host insect. , 2017, Fungal genetics and biology : FG & B.

[16]  T. Garrett,et al.  Growth substrates and caleosin-mediated functions affect conidial virulence in the insect pathogenic fungus Beauveria bassiana. , 2016, Microbiology.

[17]  A. Marty,et al.  Overexpression of Fusarium solani lipase in Pichia pastoris and its application in lipid degradation , 2016 .

[18]  M. Tanokura,et al.  Expression, purification, refolding, and enzymatic characterization of two secretory phospholipases A₂ from Neurospora crassa. , 2015, Protein expression and purification.

[19]  Y. Pei,et al.  Involvement of a caleosin in lipid storage, spore dispersal, and virulence in the entomopathogenic filamentous fungus, Beauveria bassiana. , 2015, Environmental microbiology.

[20]  Y. Pei,et al.  A novel mitochondrial membrane protein, Ohmm, limits fungal oxidative stress resistance and virulence in the insect fungal pathogen Beauveria bassiana. , 2015, Environmental microbiology.

[21]  M. Welte Expanding Roles for Lipid Droplets , 2015, Current Biology.

[22]  D. M. Mamatha,et al.  Epoxide hydrolase activities and epoxy fatty acids in the mosquito Culex quinquefasciatus. , 2015, Insect biochemistry and molecular biology.

[23]  Ananya Barman,et al.  Multiple cellular roles of Neurospora crassa plc-1, splA2, and cpe-1 in regulation of cytosolic free calcium, carotenoid accumulation, stress responses, and acquisition of thermotolerance , 2015, Journal of Microbiology.

[24]  I. Križaj Roles of secreted phospholipases A₂ in the mammalian immune system. , 2014, Protein and peptide letters.

[25]  M. Gelb,et al.  Platelets release mitochondria serving as substrate for bactericidal group IIA-secreted phospholipase A2 to promote inflammation. , 2014, Blood.

[26]  D. Stanley,et al.  Roles of Peroxinectin in PGE2-Mediated Cellular Immunity in Spodoptera exigua , 2014, PloS one.

[27]  Chengshu Wang,et al.  Advances in fundamental and applied studies in China of fungal biocontrol agents for use against arthropod pests , 2014 .

[28]  Q. Gao,et al.  Glycerol-3-Phosphate Acyltransferase Contributes to Triacylglycerol Biosynthesis, Lipid Droplet Formation, and Host Invasion in Metarhizium robertsii , 2013, Applied and Environmental Microbiology.

[29]  N. Keyhani,et al.  Action on the Surface: Entomopathogenic Fungi versus the Insect Cuticle , 2013, Insects.

[30]  Ellen C. Jensen* Quantitative Analysis of Histological Staining and Fluorescence Using ImageJ , 2013, Anatomical record.

[31]  S. Ying,et al.  Catalases play differentiated roles in the adaptation of a fungal entomopathogen to environmental stresses. , 2013, Environmental microbiology.

[32]  Shizhu Zhang,et al.  Targeting of insect epicuticular lipids by the entomopathogenic fungus Beauveria bassiana: hydrocarbon oxidation within the context of a host-pathogen interaction , 2013, Front. Microbio..

[33]  Guo-Ping Zhao,et al.  Genomic perspectives on the evolution of fungal entomopathogenicity in Beauveria bassiana , 2012, Scientific Reports.

[34]  Y. Pei,et al.  The MAP kinase Bbslt2 controls growth, conidiation, cell wall integrity, and virulence in the insect pathogenic fungus Beauveria bassiana. , 2012, Fungal genetics and biology : FG & B.

[35]  Shizhu Zhang,et al.  CYP52X1, Representing New Cytochrome P450 Subfamily, Displays Fatty Acid Hydroxylase Activity and Contributes to Virulence and Growth on Insect Cuticular Substrates in Entomopathogenic Fungus Beauveria bassiana* , 2012, The Journal of Biological Chemistry.

[36]  M. Gelb,et al.  Phospholipases: an overview. , 2012, Methods in molecular biology.

[37]  E. Dennis,et al.  Phospholipase A2 enzymes: physical structure, biological function, disease implication, chemical inhibition, and therapeutic intervention. , 2011, Chemical reviews.

[38]  Shizhu Zhang,et al.  Contribution of the gas1 Gene of the Entomopathogenic Fungus Beauveria bassiana, Encoding a Putative Glycosylphosphatidylinositol-Anchored β-1,3-Glucanosyltransferase, to Conidial Thermotolerance and Virulence , 2011, Applied and Environmental Microbiology.

[39]  S. Rivas,et al.  Phospholipases in action during plant defense signaling , 2011, Plant signaling & behavior.

[40]  H. J. Kim,et al.  Endoplasmic Reticulum– and Golgi-Localized Phospholipase A2 Plays Critical Roles in Arabidopsis Pollen Development and Germination[W][OA] , 2011, Plant Cell.

[41]  D. Roby,et al.  AtsPLA2-α nuclear relocalization by the Arabidopsis transcription factor AtMYB30 leads to repression of the plant defense response , 2010, Proceedings of the National Academy of Sciences.

[42]  Daisuke Fujioka,et al.  A novel anti-inflammatory role for secretory phospholipase A2 in immune complex-mediated arthritis , 2010, EMBO molecular medicine.

[43]  S. Ottonello,et al.  Distinct enzymatic and cellular characteristics of two secretory phospholipases A2 in the filamentous fungus Aspergillus oryzae. , 2010, Fungal genetics and biology : FG & B.

[44]  D. Stanley,et al.  Genes encoding phospholipases A2 mediate insect nodulation reactions to bacterial challenge. , 2010, Journal of insect physiology.

[45]  Y. Pei,et al.  Requirement of a Mitogen-Activated Protein Kinase for Appressorium Formation and Penetration of Insect Cuticle by the Entomopathogenic Fungus Beauveria bassiana , 2010, Applied and Environmental Microbiology.

[46]  Y. Pei,et al.  Expressing a fusion protein with protease and chitinase activities increases the virulence of the insect pathogen Beauveria bassiana. , 2009, Journal of invertebrate pathology.

[47]  N. Keyhani,et al.  Lectin mapping reveals stage-specific display of surface carbohydrates in in vitro and haemolymph-derived cells of the entomopathogenic fungus Beauveria bassiana. , 2009, Microbiology.

[48]  N. Keyhani,et al.  Uptake of the fluorescent probe FM4-64 by hyphae and haemolymph-derived in vivo hyphal bodies of the entomopathogenic fungus Beauveria bassiana. , 2009, Microbiology.

[49]  J. Napier,et al.  Misexpression of FATTY ACID ELONGATION1 in the Arabidopsis Epidermis Induces Cell Death and Suggests a Critical Role for Phospholipase A2 in This Process[W] , 2009, The Plant Cell Online.

[50]  Y. Pei,et al.  The size and ratio of homologous sequence to non-homologous sequence in gene disruption cassette influences the gene targeting efficiency in Beauveria bassiana , 2009, Applied Microbiology and Biotechnology.

[51]  J. Gardiner,et al.  The phospholipase A inhibitor, aristolochic acid, disrupts cortical microtubule arrays and root growth in Arabidopsis. , 2008, Plant biology.

[52]  A. Padhi,et al.  Detecting molecular adaptation at individual codons in the pattern recognition protein, lipopolysaccharide- and beta-1,3-glucan-binding protein of decapods. , 2008, Fish & shellfish immunology.

[53]  Xingyong Yang,et al.  A cuticle-degrading protease (CDEP-1) of Beauveria bassiana enhances virulence , 2008 .

[54]  Xingyong Yang,et al.  Expression of a Beauveria bassiana chitinase (Bbchit1) in Escherichia coli and Pichia pastoris. , 2007, Protein expression and purification.

[55]  M. Murakami,et al.  Human group III phospholipase A2 suppresses adenovirus infection into host cells. Evidence that group III, V and X phospholipase A2s act on distinct cellular phospholipid molecular species. , 2007, Biochimica et biophysica acta.

[56]  Chengshu Wang,et al.  The Metarhizium anisopliae Perilipin Homolog MPL1 Regulates Lipid Metabolism, Appressorial Turgor Pressure, and Virulence* , 2007, Journal of Biological Chemistry.

[57]  Edward A Dennis,et al.  The phospholipase A2 superfamily and its group numbering system. , 2006, Biochimica et biophysica acta.

[58]  N. Pedrini,et al.  Clues on the role of Beauveria bassiana catalases in alkane degradation events. , 2006, Mycologia.

[59]  S. Ying,et al.  Novel blastospore-based transformation system for integration of phosphinothricin resistance and green fluorescence protein genes into Beauveria bassiana , 2006, Applied Microbiology and Biotechnology.

[60]  L. Miozzi,et al.  Phospholipase A2 up-regulation during mycorrhiza formation in Tuber borchii. , 2005, The New phytologist.

[61]  I. Hwang,et al.  Multiple forms of secretory phospholipase A2 in plants. , 2005, Progress in lipid research.

[62]  B. L. Diaz,et al.  Phospholipase A(2). , 2003, Prostaglandins, leukotrienes, and essential fatty acids.

[63]  G. Lambeau,et al.  A peptide derived from bee venom-secreted phospholipase A2 inhibits replication of T-cell tropic HIV-1 strains via interaction with the CXCR4 chemokine receptor. , 2001, Molecular pharmacology.

[64]  M. Ghannoum Potential role of phospholipases in virulence and fungal pathogenesis. , 2000, Clinical microbiology reviews.