A collagenous protective coat enables Metarhizium anisopliae to evade insect immune responses.
The ubiquitous fungal pathogen Metarhizium anisopliae kills a wide range of insects. Host hemocytes can recognize and ingest its conidia, but this capacity is lost on production of hyphal bodies. We show that the unusual ability of hyphal bodies to avoid detection depends on a gene (Mcl1) that is expressed within 20 min of the pathogen contacting hemolymph. A mutant disrupted in Mcl1 is rapidly attacked by hemocytes and shows a corresponding reduction of virulence to Manduca sexta. Mcl1 encodes a three domain protein comprising a hydrophilic, negatively charged N-terminal region with 14 cysteine residues, a central region comprising tandem repeats (GXY) characteristic of collagenous domains, and a C-terminal region that includes a glycosylphosphatidylinositol-dependent cell wall attachment site. Immunofluorescence assay showed that hyphal bodies are covered by the N-terminal domains of MCL1. The collagen domain became antibody accessible after treatment with DTT, suggesting that the N termini are linked by interchain disulfide bonds and are presented on the cell surface by extended collagenous fibers. Studies with staining reagents and hemocyte monolayers showed that MCL1 functions as an antiadhesive protective coat because it masks antigenic structural components of the cell wall such as beta-glucans, and because its hydrophilic negatively charged nature makes it unattractive to hemocytes. A survey of 54 fungal genomes revealed that seven other species have proteins with collagenous domains suggesting that MCL1 is a member of a patchily distributed gene family.
Wang
Chengshu
C
Department of Entomology, University of Maryland, College Park, MD 20742, USA.
St Leger
Raymond J
RJ
D000595:Amino Acid Sequence; D000818:Animals; D003094:Collagen / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D018390:Gene Targeting; D005800:Genes, Fungal; D006434:Hemocytes / Q000276:immunology / Q000382:microbiology; D006458:Hemolymph / Q000276:immunology / Q000382:microbiology; D006790:Host-Parasite Interactions; D006999:Hypocreales / Q000235:genetics / Q000276:immunology / Q000378:metabolism* / Q000472:pathogenicity*; D066298:In Vitro Techniques; D007313:Insecta / Q000276:immunology* / Q000382:microbiology*; D008969:Molecular Sequence Data; D009154:Mutation; D011499:Protein Processing, Post-Translational; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D014774:Virulence / Q000235:genetics / Q000502:physiology
D005656:Fungal Proteins; D003094:Collagen