A small molecule species specifically inhibits Fusarium myosin I
Chengqi Zhang
Institute of Biotechnology, Zhejiang University, Hangzhou, 310058 China
These authors contributed equally to this work.Search for more papers by this authorYun Chen
Institute of Biotechnology, Zhejiang University, Hangzhou, 310058 China
These authors contributed equally to this work.Search for more papers by this authorYanni Yin
Institute of Biotechnology, Zhejiang University, Hangzhou, 310058 China
Search for more papers by this authorHuan-Hong Ji
National Laboratory of Integrated Management of Insect Pests and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101 China
Search for more papers by this authorWon-Bo Shim
Department of Plant Pathology and Microbiology, Texas A&M University, College Station, TX, 77843-2132 USA
Search for more papers by this authorYiping Hou
Department of Plant Pathology, Nanjing Agricultural University, Nanjing, 210095 China
Search for more papers by this authorCorresponding Author
Mingguo Zhou
Department of Plant Pathology, Nanjing Agricultural University, Nanjing, 210095 China
For correspondence. E-mail [email protected]; [email protected] or [email protected]; Tel. (+86) 571 88982268; Fax (+86) 571 88982268.Search for more papers by this authorCorresponding Author
Xiang-dong Li
National Laboratory of Integrated Management of Insect Pests and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101 China
For correspondence. E-mail [email protected]; [email protected] or [email protected]; Tel. (+86) 571 88982268; Fax (+86) 571 88982268.Search for more papers by this authorCorresponding Author
Zhonghua Ma
Institute of Biotechnology, Zhejiang University, Hangzhou, 310058 China
For correspondence. E-mail [email protected]; [email protected] or [email protected]; Tel. (+86) 571 88982268; Fax (+86) 571 88982268.Search for more papers by this authorChengqi Zhang
Institute of Biotechnology, Zhejiang University, Hangzhou, 310058 China
These authors contributed equally to this work.Search for more papers by this authorYun Chen
Institute of Biotechnology, Zhejiang University, Hangzhou, 310058 China
These authors contributed equally to this work.Search for more papers by this authorYanni Yin
Institute of Biotechnology, Zhejiang University, Hangzhou, 310058 China
Search for more papers by this authorHuan-Hong Ji
National Laboratory of Integrated Management of Insect Pests and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101 China
Search for more papers by this authorWon-Bo Shim
Department of Plant Pathology and Microbiology, Texas A&M University, College Station, TX, 77843-2132 USA
Search for more papers by this authorYiping Hou
Department of Plant Pathology, Nanjing Agricultural University, Nanjing, 210095 China
Search for more papers by this authorCorresponding Author
Mingguo Zhou
Department of Plant Pathology, Nanjing Agricultural University, Nanjing, 210095 China
For correspondence. E-mail [email protected]; [email protected] or [email protected]; Tel. (+86) 571 88982268; Fax (+86) 571 88982268.Search for more papers by this authorCorresponding Author
Xiang-dong Li
National Laboratory of Integrated Management of Insect Pests and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101 China
For correspondence. E-mail [email protected]; [email protected] or [email protected]; Tel. (+86) 571 88982268; Fax (+86) 571 88982268.Search for more papers by this authorCorresponding Author
Zhonghua Ma
Institute of Biotechnology, Zhejiang University, Hangzhou, 310058 China
For correspondence. E-mail [email protected]; [email protected] or [email protected]; Tel. (+86) 571 88982268; Fax (+86) 571 88982268.Search for more papers by this authorSummary
Fusarium head blight (FHB) caused by Fusarium graminearum is a devastating disease of cereal crops worldwide. Recently, a novel fungicide JS399-19 has been launched into the marketplace to manage FHB. It is compelling that JS399-19 shows highly inhibitory activity towards some Fusarium species, but not to other fungi, indicating that it is an environmentally compatible fungicide. To explore the mode of action of this species-specific compound, we conducted a whole-genome transcript profiling together with genetic and biochemical assays, and discovered that JS399-19 targets the myosin I of F. graminearum (FgMyo1). FgMyo1 is essential for F. graminearum growth. A point mutation S217L or E420K in FgMyo1 is responsible for F. graminearum resistance to JS399-19. In addition, transformation of F. graminearum with the myosin I gene of Magnaporthe grisea, the causal agent of rice blast, also led to JS399-19 resistance. JS399-19 strongly inhibits the ATPase activity of the wild-type FgMyo1, but not the mutated FgMyo1S217L/E420K. These results provide us a new insight into the design of species-specific antifungal compounds. Furthermore, our strategy can be applied to identify novel drug targets in various pathogenic organisms.
Supporting Information
| Filename | Description |
|---|---|
| emi12711-sup-0001-si.tif1.8 MB | Fig. S1. Inhibitory effect of carbendazim, tebuconazole and JS399-19 against F. graminearum and F. asiaticum. A. Chemical structure of the novel fungicide JS399-19. B. Fusarium graminearum strain PH-1 and F. asiaticum strain GJ33 were inoculated onto the PDA plates supplemented with 0.3 μg ml−1 tested fungicides. The solvent DMSO was used as a control. Inoculated plates were photographed after 3 days incubation at 25°C. |
| emi12711-sup-0002-si.tif4.1 MB | Fig. S2. Inhibitory effect of JS399-19 against conidial germination of F. graminearum and Magnaporthe grisea after 2, 4 or 8 h of incubation. |
| emi12711-sup-0003-si.tif94.6 KB | Fig. S3. The transcription level of the beta-tubulin gene (TUB2) in F. graminearum PH-1 treated with carbendazim. The relative transcription level of TUB2 in PH-1 treated with 10 μg ml−1 carbendazim is the relative amount of mRNA of the gene in PH-1 without carbendazim treatment (CK). |
| emi12711-sup-0004-si.tif2 MB | Fig. S4. JS399-19-resistant (JS-R) mutants obtained by UV irradiation contain the point mutations at codon 217 and 420 of FgMyo1. A. The mutants (JS-R-1 to 10) and the wild-type PH-1 were incubated on PDA amended with 1 or 100 μg ml−1 JS399-19. The solvent DMSO was used as a control. B. Alignment of partial deduced amino acid sequences of FgMyo1 from the wild-type PH-1 and JS-R mutants. The vertical boxes indicate the amino acid changes at the codons 217 and 420 that are responsible for JS399-19 resistance. C. Predicted domains of FgMyo1 protein. Line represents the full-length 1129-amino-acid protein. The boxes represent the identified domains and were labelled with different colors. D. Locations of point mutations S217L and E420K in the putative FgMyo1 structure, which was constructed based on the crystal structure of Dictyostelium discoideum myosin IE motor domain (PDB code 1LKX). Green, 25 kDa domain; red, upper 50 kDa domain; white, lower 50 kDa domain, blue, 20 kDa domain; spheres, Mg2+.ADP.VO4. Structure model of FgMyo1 was constructed using the PyMOL Molecular Graphics System software (DeLano Scientific, San Carlos, CA, USA). |
| emi12711-sup-0005-si.tif2.1 MB | Fig. S5. Southern blotting assays of FgMyo1 in the wild-type PH-1 and the strains derived from PH-1. |
| emi12711-sup-0006-si.tif14.5 MB | Fig. S6. Alignments of amino acid sequences of Myo1 from different fungi, including Fusarium graminearum (FgMyo1), Fusarium verticillioides (FvMyo1), Fusarium oxysporum (FoMyo1), Fusarium solani (FsMyo1), Magnaporthe grisea (MgMyo1), Botrytis cinerea (BcMyo1), Aspergillus flavus (AfMyo1) and Saccharomyces cerevisiae (ScMyo1). The vertical blue and red boxes indicate the amino acid residues corresponding to the S217 and E420 of FgMyo1 respectively. |
| emi12711-sup-0007-si.tif459.8 KB | Fig. S7. SDS-PAGE (4–20%) of the purified FgMyo1-MIQWT, FgMyo1-MIQS217L and FgMyo1-MIQE420K. The arrowhead indicates the target protein. |
| emi12711-sup-0008-si.docx23 KB | Table S1. A list of 156 genes upregulated by JS399-19 treatment in the wild-type PH-1. |
| emi12711-sup-0009-si.doc806.5 KB | Table S2. PCR primers used in this study. |
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