Related Experiment Video
Updated: Dec 14, 2025
Genome-wide Analysis of Histone Modifications Distribution using the Chromatin Immunoprecipitation Sequencing Method in Magnaporthe oryzae
Published on: June 2, 2021
Comparative Proteomic Analysis of Wild Type and Mutant Lacking an SCF E3 Ligase F-Box Protein in Magnaporthe oryzae
Abstract:
Magnaporthe oryzae (M. oryzae) is a pathogenic, filamentous fungus that is a primary cause of rice blast disease. The M. oryzae protein MGG_13065, SCF E3 ubiquitin ligase complex F-box protein, has been identified as playing a crucial role in the infection process, specifically, as part of the ubiquitin mediated proteolysis pathway. Proteins targeted by MGG_13065 E3 ligase are first phosphorylated and then ubiquitinated by E3 ligase. In this study, we used a label-free quantitative global proteomics technique to probe the role of ubiquitination and phosphorylation in the mechanism of how E3 ligase regulates change in virulence of M. oryzae. To do this, we compared the WT M. oryzae 70-15 strain with a gene knock out (E3 ligase KO) strain. After applying a ≥ 5 normalized spectral count cutoff, a total of 4432 unique proteins were identified comprised of 4360 and 4372 in the WT and E3 ligase KO samples, respectively. Eighty proteins drastically increased in abundance, while 65 proteins decreased in abundance in the E3 ligase KO strain. Proteins (59) were identified only in the WT strain; 13 of these proteins had both phosphorylation and ubiquitination post-translational modifications. Proteins (71) were revealed to be only in the E3 ligase KO strain; 23 of the proteins have both phosphorylation and ubiquitination post-translational modifications. Several of these proteins were associated with key biological processes. These data greatly assist in the selection of future genes for functional studies and enable mechanistic insight related to virulence.
Insights
This study investigates how the Magnaporthe oryzae E3 ligase regulates rice blast virulence using proteomics. It reveals key proteins and post-translational modifications involved in the infection process.
Area of Science:
- Plant Pathology
- Molecular Biology
- Fungal Proteomics
Background:
- Magnaporthe oryzae causes rice blast disease.
- The MGG_13065 protein, an SCF E3 ubiquitin ligase complex F-box protein, is vital for M. oryzae infection.
- Ubiquitination and phosphorylation are key post-translational modifications in protein regulation.
Purpose of the Study:
- To investigate the role of ubiquitination and phosphorylation in E3 ligase-mediated virulence regulation in M. oryzae.
- To compare protein abundance and post-translational modifications between wild-type (WT) and E3 ligase knockout (KO) M. oryzae strains.
- To identify proteins and biological processes affected by E3 ligase activity.
Main Methods:
- Label-free quantitative global proteomics.
- Comparative analysis of WT 70-15 and E3 ligase KO M. oryzae strains.
- Identification and analysis of differentially abundant proteins and post-translational modifications (ubiquitination and phosphorylation).
Main Results:
- 4432 unique proteins identified in total.
- Significant changes in protein abundance observed in the E3 ligase KO strain (80 increased, 65 decreased).
- Specific proteins with dual phosphorylation and ubiquitination modifications were identified uniquely in either WT or E3 ligase KO strains, suggesting their role in virulence regulation.
Conclusions:
- The study provides mechanistic insights into how E3 ligase regulates M. oryzae virulence through protein ubiquitination and phosphorylation.
- Identified proteins and pathways offer targets for future functional studies to combat rice blast disease.
- Proteomics data significantly advance understanding of M. oryzae pathogenesis and virulence factors.
