Journal
JOURNAL OF PROTEOME RESEARCH
Volume 15, Issue 3, Pages 826-839Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.5b00732
Keywords
biotrophic pathogen; barley; Blumeria graminis; RNase-like effector; elongation factor; pathogenesis-related protein-S; glutathione-S-transferase (GST); malate dehydrogenase
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Funding
- BBSRC [BB/H001646/1, BB/H001948/1]
- BBSRC Doctoral Training Grant [BB/F017324/1]
- BBSRC [BB/H001646/1, BB/H001948/1, BB/M000710/1, BB/E000983/1] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/H001948/1, 1103887, BB/E000983/1, BB/H001646/1, BB/C513218/1, BB/M000710/1] Funding Source: researchfish
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There are over 500 candidate secreted effector proteins (CSEPs) or Blumeria effector candidates (BECs) specific to the barley powdery mildew pathogen Blumeria graminis fsp. hordei. The CSEP/BEC proteins are expressed and predicted to be secreted by biotrophic feeding structures called haustoria. Eight BECs are required for the formation of functional haustoria. These include the RNase-like effector BEC1054 (synonym CSEP0064). In order to identify host proteins targeted by BEC1054, recombinant BEC1054 was expressed in E. coli, solubilized, and used in pull-down assays from barley protein extracts. Many putative interactors were identified by LC-MS/MS after subtraction of unspecific binders in negative controls. Therefore, a directed yeast-2 hybrid assay, developed to measure the effectiveness of the interactions in yeast, was used to validate putative interactors. We conclude that BEC1054 may target several host proteins, including a glutathione-S-transferase, a malate dehydrogenase, and a pathogen-related-5 protein isoform, indicating a possible role for BEC1054 in compromising well-known key players of defense and response to pathogens. In addition, BEC1054 interacts with an elongation factor 1 gamma. This study already suggests that BEC1054 plays a central role in barley powdery mildew virulence by acting at several levels.
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