Interactions of Destruxin A with Silkworms’ Arginine tRNA Synthetase and Lamin-C Proteins
Destruxin A (DA), a cyclodepsipeptidic mycotoxin produced by entomopathogenic fungus <i>Metarhizium anisopliae</i>, has good insecticidal activity and potential to be a new pesticide. However, the mechanism of action is still obscure. Our previous experiments showed that DA was involved...
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| Format: | Article |
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MDPI AG
2020-02-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/12/2/137 |
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| author | Jingjing Wang Qunfang Weng Fei Yin Qiongbo Hu |
| author_facet | Jingjing Wang Qunfang Weng Fei Yin Qiongbo Hu |
| author_sort | Jingjing Wang |
| collection | DOAJ |
| description | Destruxin A (DA), a cyclodepsipeptidic mycotoxin produced by entomopathogenic fungus <i>Metarhizium anisopliae</i>, has good insecticidal activity and potential to be a new pesticide. However, the mechanism of action is still obscure. Our previous experiments showed that DA was involved in regulation of transcription and protein synthesis and suggested that silkworms’ arginine tRNA synthetase (BmArgRS), Lamin-C Proteins (BmLamin-C) and ATP-dependent RNA helicase PRP1 (BmPRP1) were candidates of DA-binding proteins. In this study, we employed bio-layer interferometry (BLI), circular dichroism (CD), cellular thermal shift assay (CETSA), and other technologies to verify the interaction of DA with above three proteins in vitro and in vivo. The results of BLI indicated that BmArgRS and BmLamin-C were binding-protein of DA with K<sub>D</sub> value 5.53 × 10<sup>−5</sup> and 8.64 × 10<sup>−5</sup> M, but not BmPRP1. These interactions were also verified by CD and CETSA tests. In addition, docking model and mutants assay in vitro showed that BmArgRS interacts with DA at the pocket including Lys228, His231, Asp434 and Gln437 in its enzyme active catalysis region, while BmLamin-C binds to DA at His524 and Lys528 in the tail domain. This study might provide new insight and evidence in illustrating molecular mechanism of DA in breaking insect. |
| first_indexed | 2024-04-13T08:26:36Z |
| format | Article |
| id | doaj.art-35cf42e53bb64304862af2dad91d8eef |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-13T08:26:36Z |
| publishDate | 2020-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-35cf42e53bb64304862af2dad91d8eef2022-12-22T02:54:26ZengMDPI AGToxins2072-66512020-02-0112213710.3390/toxins12020137toxins12020137Interactions of Destruxin A with Silkworms’ Arginine tRNA Synthetase and Lamin-C ProteinsJingjing WangQunfang WengFei YinQiongbo HuDestruxin A (DA), a cyclodepsipeptidic mycotoxin produced by entomopathogenic fungus <i>Metarhizium anisopliae</i>, has good insecticidal activity and potential to be a new pesticide. However, the mechanism of action is still obscure. Our previous experiments showed that DA was involved in regulation of transcription and protein synthesis and suggested that silkworms’ arginine tRNA synthetase (BmArgRS), Lamin-C Proteins (BmLamin-C) and ATP-dependent RNA helicase PRP1 (BmPRP1) were candidates of DA-binding proteins. In this study, we employed bio-layer interferometry (BLI), circular dichroism (CD), cellular thermal shift assay (CETSA), and other technologies to verify the interaction of DA with above three proteins in vitro and in vivo. The results of BLI indicated that BmArgRS and BmLamin-C were binding-protein of DA with K<sub>D</sub> value 5.53 × 10<sup>−5</sup> and 8.64 × 10<sup>−5</sup> M, but not BmPRP1. These interactions were also verified by CD and CETSA tests. In addition, docking model and mutants assay in vitro showed that BmArgRS interacts with DA at the pocket including Lys228, His231, Asp434 and Gln437 in its enzyme active catalysis region, while BmLamin-C binds to DA at His524 and Lys528 in the tail domain. This study might provide new insight and evidence in illustrating molecular mechanism of DA in breaking insect.https://www.mdpi.com/2072-6651/12/2/137destruxinsbombyx moribmargrsbmlamin-crna helicasebinding protein |
| spellingShingle | Jingjing Wang Qunfang Weng Fei Yin Qiongbo Hu Interactions of Destruxin A with Silkworms’ Arginine tRNA Synthetase and Lamin-C Proteins Toxins destruxins bombyx mori bmargrs bmlamin-c rna helicase binding protein |
| title | Interactions of Destruxin A with Silkworms’ Arginine tRNA Synthetase and Lamin-C Proteins |
| title_full | Interactions of Destruxin A with Silkworms’ Arginine tRNA Synthetase and Lamin-C Proteins |
| title_fullStr | Interactions of Destruxin A with Silkworms’ Arginine tRNA Synthetase and Lamin-C Proteins |
| title_full_unstemmed | Interactions of Destruxin A with Silkworms’ Arginine tRNA Synthetase and Lamin-C Proteins |
| title_short | Interactions of Destruxin A with Silkworms’ Arginine tRNA Synthetase and Lamin-C Proteins |
| title_sort | interactions of destruxin a with silkworms arginine trna synthetase and lamin c proteins |
| topic | destruxins bombyx mori bmargrs bmlamin-c rna helicase binding protein |
| url | https://www.mdpi.com/2072-6651/12/2/137 |
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