Identification and characterization of the host protein DNAJC14 as a broadly active flavivirus replication modulator.
Viruses in the Flavivirus genus of the Flaviviridae family are arthropod-transmitted and contribute to staggering numbers of human infections and significant deaths annually across the globe. To identify cellular factors with antiviral activity against flaviviruses, we screened a cDNA library using...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS Pathogens |
| Online Access: | https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1001255&type=printable |
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| author | Zhigang Yi Lindsey Sperzel Cindy Nürnberger Peter J Bredenbeek Kirk J Lubick Sonja M Best Cristina T Stoyanov Lok Man J Law Zhenghong Yuan Charles M Rice Margaret R MacDonald |
| author_facet | Zhigang Yi Lindsey Sperzel Cindy Nürnberger Peter J Bredenbeek Kirk J Lubick Sonja M Best Cristina T Stoyanov Lok Man J Law Zhenghong Yuan Charles M Rice Margaret R MacDonald |
| author_sort | Zhigang Yi |
| collection | DOAJ |
| description | Viruses in the Flavivirus genus of the Flaviviridae family are arthropod-transmitted and contribute to staggering numbers of human infections and significant deaths annually across the globe. To identify cellular factors with antiviral activity against flaviviruses, we screened a cDNA library using an iterative approach. We identified a mammalian Hsp40 chaperone protein (DNAJC14) that when overexpressed was able to mediate protection from yellow fever virus (YFV)-induced cell death. Further studies revealed that DNAJC14 inhibits YFV at the step of viral RNA replication. Since replication of bovine viral diarrhea virus (BVDV), a member of the related Pestivirus genus, is also known to be modulated by DNAJC14, we tested the effect of this host factor on diverse Flaviviridae family members. Flaviviruses, including the pathogenic Asibi strain of YFV, Kunjin, and tick-borne Langat virus, as well as a Hepacivirus, hepatitis C virus (HCV), all were inhibited by overexpression of DNAJC14. Mutagenesis showed that both the J-domain and the C-terminal domain, which mediates self-interaction, are required for anti-YFV activity. We found that DNAJC14 does not block YFV nor HCV NS2-3 cleavage, and using non-inhibitory mutants demonstrate that DNAJC14 is recruited to YFV replication complexes. Immunofluorescence analysis demonstrated that endogenous DNAJC14 rearranges during infection and is found in replication complexes identified by dsRNA staining. Interestingly, silencing of endogenous DNAJC14 results in impaired YFV replication suggesting a requirement for DNAJC14 in YFV replication complex assembly. Finally, the antiviral activity of overexpressed DNAJC14 occurs in a time- and dose-dependent manner. DNAJC14 overexpression may disrupt the proper stoichiometry resulting in inhibition, which can be overcome upon restoration of the optimal ratios due to the accumulation of viral nonstructural proteins. Our findings, together with previously published work, suggest that the members of the Flaviviridae family have evolved in unique and important ways to interact with this host Hsp40 chaperone molecule. |
| format | Article |
| id | doaj-art-bff01d06ed4c4506a6daef1243d5de93 |
| institution | Kabale University |
| issn | 1553-7366 1553-7374 |
| language | English |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj-art-bff01d06ed4c4506a6daef1243d5de932025-08-20T03:47:04ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742011-01-0171e100125510.1371/journal.ppat.1001255Identification and characterization of the host protein DNAJC14 as a broadly active flavivirus replication modulator.Zhigang YiLindsey SperzelCindy NürnbergerPeter J BredenbeekKirk J LubickSonja M BestCristina T StoyanovLok Man J LawZhenghong YuanCharles M RiceMargaret R MacDonaldViruses in the Flavivirus genus of the Flaviviridae family are arthropod-transmitted and contribute to staggering numbers of human infections and significant deaths annually across the globe. To identify cellular factors with antiviral activity against flaviviruses, we screened a cDNA library using an iterative approach. We identified a mammalian Hsp40 chaperone protein (DNAJC14) that when overexpressed was able to mediate protection from yellow fever virus (YFV)-induced cell death. Further studies revealed that DNAJC14 inhibits YFV at the step of viral RNA replication. Since replication of bovine viral diarrhea virus (BVDV), a member of the related Pestivirus genus, is also known to be modulated by DNAJC14, we tested the effect of this host factor on diverse Flaviviridae family members. Flaviviruses, including the pathogenic Asibi strain of YFV, Kunjin, and tick-borne Langat virus, as well as a Hepacivirus, hepatitis C virus (HCV), all were inhibited by overexpression of DNAJC14. Mutagenesis showed that both the J-domain and the C-terminal domain, which mediates self-interaction, are required for anti-YFV activity. We found that DNAJC14 does not block YFV nor HCV NS2-3 cleavage, and using non-inhibitory mutants demonstrate that DNAJC14 is recruited to YFV replication complexes. Immunofluorescence analysis demonstrated that endogenous DNAJC14 rearranges during infection and is found in replication complexes identified by dsRNA staining. Interestingly, silencing of endogenous DNAJC14 results in impaired YFV replication suggesting a requirement for DNAJC14 in YFV replication complex assembly. Finally, the antiviral activity of overexpressed DNAJC14 occurs in a time- and dose-dependent manner. DNAJC14 overexpression may disrupt the proper stoichiometry resulting in inhibition, which can be overcome upon restoration of the optimal ratios due to the accumulation of viral nonstructural proteins. Our findings, together with previously published work, suggest that the members of the Flaviviridae family have evolved in unique and important ways to interact with this host Hsp40 chaperone molecule.https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1001255&type=printable |
| spellingShingle | Zhigang Yi Lindsey Sperzel Cindy Nürnberger Peter J Bredenbeek Kirk J Lubick Sonja M Best Cristina T Stoyanov Lok Man J Law Zhenghong Yuan Charles M Rice Margaret R MacDonald Identification and characterization of the host protein DNAJC14 as a broadly active flavivirus replication modulator. PLoS Pathogens |
| title | Identification and characterization of the host protein DNAJC14 as a broadly active flavivirus replication modulator. |
| title_full | Identification and characterization of the host protein DNAJC14 as a broadly active flavivirus replication modulator. |
| title_fullStr | Identification and characterization of the host protein DNAJC14 as a broadly active flavivirus replication modulator. |
| title_full_unstemmed | Identification and characterization of the host protein DNAJC14 as a broadly active flavivirus replication modulator. |
| title_short | Identification and characterization of the host protein DNAJC14 as a broadly active flavivirus replication modulator. |
| title_sort | identification and characterization of the host protein dnajc14 as a broadly active flavivirus replication modulator |
| url | https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1001255&type=printable |
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