Protein polyubiquitination plays a role in basal host resistance of barley.
To study protein ubiquitination pathways in the interaction of barley (Hordeum vulgare) with the powdery mildew fungus (Blumeria graminis), we measured protein turnover and performed transient-induced gene silencing (TIGS) of ubiquitin and 26S proteasome subunit encoding genes in epidermal cells. Attack by B. graminis hyperdestabilized a novel unstable green fluorescent protein fusion that contains a destabilization domain of a putative barley 1-aminocyclopropane-1-carboxylate synthase, suggesting enhanced protein turnover. Partial depletion of cellular ubiquitin levels by TIGS induced extreme susceptibility of transformed cells toward the appropriate host pathogen B. graminis f. sp hordei, whereas papilla-based resistance to the nonhost pathogen B. graminis f. sp tritici and host resistance mediated by the mlo gene (for mildew resistance locus O) remained unaffected. Cells were rescued from TIGS-induced ubiquitin depletion by synthetic genes encoding wild-type or mutant barley monoubiquitin proteins. The strongest rescue was from a gene encoding a K63R mutant form of ubiquitin blocked in several ubiquitination pathways while still allowing Lys-48-dependent polyubiquitination required for proteasomal protein degradation. Systematic RNA interference of 40 genes encoding all 17 subunits of the proteasome 19S regulatory particle failed to induce hypersusceptibility against B. graminis f. sp hordei. This suggests a role for Lys-48-linked protein polyubiquitination, which is independent from the proteasome pathway, in basal host defense of barley.
Dong
Wubei
W
Leibniz-Institute of Plant Genetics and Crop Plant Research, D-06466 Gatersleben, Germany.
Nowara
Daniela
D
Schweizer
Patrick
P
D001203:Ascomycota / Q000472:pathogenicity; D015870:Gene Expression; D020868:Gene Silencing; D017343:Genes, Plant; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000378:metabolism; D001467:Hordeum / Q000235:genetics / Q000276:immunology / Q000378:metabolism* / Q000382:microbiology*; D007113:Immunity, Innate / Q000276:immunology*; D008190:Lyases / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D008954:Models, Biological; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000276:immunology*; D019441:Plant Epidermis / Q000166:cytology / Q000382:microbiology; D018515:Plant Leaves / Q000382:microbiology; D010940:Plant Proteins / Q000378:metabolism*; D025821:Polyubiquitin / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013997:Time Factors; D014908:Triticum / Q000235:genetics / Q000276:immunology* / Q000382:microbiology
D010940:Plant Proteins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D025821:Polyubiquitin; D049452:Green Fluorescent Proteins; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease; D008190:Lyases; C021773:1-aminocyclopropanecarboxylate synthase; D008239:Lysine