eEF1A Interacts with the NS5A Protein and Inhibits the Growth of Classical Swine Fever Virus
The NS5A protein of classical swine fever virus (CSFV) is involved in the RNA synthesis and viral replication. However, the NS5A-interacting cellular proteins engaged in the CSFV replication are poorly defined. Using yeast two-hybrid screen, the eukaryotic elongation factor 1A (eEF1A) was identified...
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| Format: | Article |
| Language: | English |
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MDPI AG
2015-08-01
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| Series: | Viruses |
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| Online Access: | http://www.mdpi.com/1999-4915/7/8/2833 |
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| author | Su Li Shuo Feng Jing-Han Wang Wen-Rui He Hua-Yang Qin Hong Dong Lian-Feng Li Shao-Xiong Yu Yongfeng Li Hua-Ji Qiu |
| author_facet | Su Li Shuo Feng Jing-Han Wang Wen-Rui He Hua-Yang Qin Hong Dong Lian-Feng Li Shao-Xiong Yu Yongfeng Li Hua-Ji Qiu |
| author_sort | Su Li |
| collection | DOAJ |
| description | The NS5A protein of classical swine fever virus (CSFV) is involved in the RNA synthesis and viral replication. However, the NS5A-interacting cellular proteins engaged in the CSFV replication are poorly defined. Using yeast two-hybrid screen, the eukaryotic elongation factor 1A (eEF1A) was identified to be an NS5A-binding partner. The NS5A–eEF1A interaction was confirmed by coimmunoprecipitation, glutathione S-transferase (GST) pulldown and laser confocal microscopy assays. The domain I of eEF1A was shown to be critical for the NS5A–eEF1A interaction. Overexpression of eEF1A suppressed the CSFV growth markedly, and conversely, knockdown of eEF1A enhanced the CSFV replication significantly. Furthermore, eEF1A, as well as NS5A, was found to reduce the translation efficiency of the internal ribosome entry site (IRES) of CSFV in a dose-dependent manner, as demonstrated by luciferase reporter assay. Streptavidin pulldown assay revealed that eEF1A could bind to the CSFV IRES. Collectively, our results suggest that eEF1A interacts with NS5A and negatively regulates the growth of CSFV. |
| first_indexed | 2024-12-16T08:34:06Z |
| format | Article |
| id | doaj.art-1776a0fdd1db42e7801e9cd60b3d4d66 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-12-16T08:34:06Z |
| publishDate | 2015-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-1776a0fdd1db42e7801e9cd60b3d4d662022-12-21T22:37:49ZengMDPI AGViruses1999-49152015-08-01784563458110.3390/v7082833v7082833eEF1A Interacts with the NS5A Protein and Inhibits the Growth of Classical Swine Fever VirusSu Li0Shuo Feng1Jing-Han Wang2Wen-Rui He3Hua-Yang Qin4Hong Dong5Lian-Feng Li6Shao-Xiong Yu7Yongfeng Li8Hua-Ji Qiu9State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, Heilongjiang, ChinaThe NS5A protein of classical swine fever virus (CSFV) is involved in the RNA synthesis and viral replication. However, the NS5A-interacting cellular proteins engaged in the CSFV replication are poorly defined. Using yeast two-hybrid screen, the eukaryotic elongation factor 1A (eEF1A) was identified to be an NS5A-binding partner. The NS5A–eEF1A interaction was confirmed by coimmunoprecipitation, glutathione S-transferase (GST) pulldown and laser confocal microscopy assays. The domain I of eEF1A was shown to be critical for the NS5A–eEF1A interaction. Overexpression of eEF1A suppressed the CSFV growth markedly, and conversely, knockdown of eEF1A enhanced the CSFV replication significantly. Furthermore, eEF1A, as well as NS5A, was found to reduce the translation efficiency of the internal ribosome entry site (IRES) of CSFV in a dose-dependent manner, as demonstrated by luciferase reporter assay. Streptavidin pulldown assay revealed that eEF1A could bind to the CSFV IRES. Collectively, our results suggest that eEF1A interacts with NS5A and negatively regulates the growth of CSFV.http://www.mdpi.com/1999-4915/7/8/2833classical swine fever virusvirus–host interactionsNS5Aeukaryotic elongation factor 1Ainternal ribosome entry site |
| spellingShingle | Su Li Shuo Feng Jing-Han Wang Wen-Rui He Hua-Yang Qin Hong Dong Lian-Feng Li Shao-Xiong Yu Yongfeng Li Hua-Ji Qiu eEF1A Interacts with the NS5A Protein and Inhibits the Growth of Classical Swine Fever Virus Viruses classical swine fever virus virus–host interactions NS5A eukaryotic elongation factor 1A internal ribosome entry site |
| title | eEF1A Interacts with the NS5A Protein and Inhibits the Growth of Classical Swine Fever Virus |
| title_full | eEF1A Interacts with the NS5A Protein and Inhibits the Growth of Classical Swine Fever Virus |
| title_fullStr | eEF1A Interacts with the NS5A Protein and Inhibits the Growth of Classical Swine Fever Virus |
| title_full_unstemmed | eEF1A Interacts with the NS5A Protein and Inhibits the Growth of Classical Swine Fever Virus |
| title_short | eEF1A Interacts with the NS5A Protein and Inhibits the Growth of Classical Swine Fever Virus |
| title_sort | eef1a interacts with the ns5a protein and inhibits the growth of classical swine fever virus |
| topic | classical swine fever virus virus–host interactions NS5A eukaryotic elongation factor 1A internal ribosome entry site |
| url | http://www.mdpi.com/1999-4915/7/8/2833 |
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