Interactions of the Nipah Virus P, V, and W Proteins across the STAT Family of Transcription Factors
ABSTRACT The Nipah virus (NiV) phosphoprotein (P) gene encodes four proteins. Three of these—P, V, and W—possess a common N-terminal domain but distinct C termini. These proteins interact with immune modulators. Previous studies demonstrated that P, V, and W bind STAT1 and STAT4 and that V also inte...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2020-12-01
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| Series: | mSphere |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mSphere.00449-20 |
| _version_ | 1831667549410951168 |
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| author | Timothy R. Keiffer Michael J. Ciancanelli Megan R. Edwards Christopher F. Basler |
| author_facet | Timothy R. Keiffer Michael J. Ciancanelli Megan R. Edwards Christopher F. Basler |
| author_sort | Timothy R. Keiffer |
| collection | DOAJ |
| description | ABSTRACT The Nipah virus (NiV) phosphoprotein (P) gene encodes four proteins. Three of these—P, V, and W—possess a common N-terminal domain but distinct C termini. These proteins interact with immune modulators. Previous studies demonstrated that P, V, and W bind STAT1 and STAT4 and that V also interacts with STAT2 but not with STAT3. The STAT1 and STAT2 interactions block interferon (IFN)-induced STAT tyrosine phosphorylation. To more fully characterize the interactions of P, V, and W with the STATs, we screened for interaction of each viral protein with STATs 1 to 6 by coimmunoprecipitation. We demonstrate that NiV P, V, and W interact with STAT4 through their common N-terminal domain and block STAT4 activity, based on a STAT4 response element reporter assay. Although none of the NiV proteins interact with STAT3 or STAT6, NiV V, but not P or W, interacts with STAT5 through its unique C terminus. Furthermore, the interaction of NiV V with STAT5 was not disrupted by overexpression of the N-terminal binding STAT1 or the C-terminal binding MDA5. NiV V also inhibits a STAT5 response element reporter assay. Residues 114 to 140 of the common N-terminal domain of the NiV P gene products were found to be sufficient to bind STAT1 and STAT4. Analysis of STAT1-STAT3 chimeras suggests that the P gene products target the STAT1 SH2 domain. When fused to GST, the 114-140 peptide is sufficient to decrease STAT1 phosphorylation in IFN-β-stimulated cells, suggesting that this peptide could potentially be fused to heterologous proteins to confer inhibition of STAT1- and STAT4-dependent responses. IMPORTANCE How Nipah virus (NiV) antagonizes innate immune responses is incompletely understood. The P gene of NiV encodes the P, V, and W proteins. These proteins have a common N-terminal sequence that is sufficient to bind to STAT1 and STAT2 and block IFN-induced signal transduction. This study sought to more fully understand how P, V, and W engage with the STAT family of transcription factors to influence their functions. The results identify a novel interaction of V with STAT5 and demonstrate V inhibition of STAT5 function. We also demonstrate that the common N-terminal residues 114 to 140 of P, V, and W are critical for inhibition of STAT1 and STAT4 function, map the interaction to the SH2 region of STAT1, and show that a fusion construct with this peptide significantly inhibits cytokine-induced STAT1 phosphorylation. These data clarify how these important virulence factors modulate innate antiviral defenses. |
| first_indexed | 2024-12-19T20:13:39Z |
| format | Article |
| id | doaj.art-3ec80e3a593242a58ec882a593caba86 |
| institution | Directory Open Access Journal |
| issn | 2379-5042 |
| language | English |
| last_indexed | 2024-12-19T20:13:39Z |
| publishDate | 2020-12-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mSphere |
| spelling | doaj.art-3ec80e3a593242a58ec882a593caba862022-12-21T20:07:14ZengAmerican Society for MicrobiologymSphere2379-50422020-12-015610.1128/mSphere.00449-20Interactions of the Nipah Virus P, V, and W Proteins across the STAT Family of Transcription FactorsTimothy R. Keiffer0Michael J. Ciancanelli1Megan R. Edwards2Christopher F. Basler3Center for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, Georgia, USADepartment of Microbiology, Icahn School of Medicine at Mount Sinai, New York, New York, USACenter for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, Georgia, USACenter for Microbial Pathogenesis, Institute for Biomedical Sciences, Georgia State University, Atlanta, Georgia, USAABSTRACT The Nipah virus (NiV) phosphoprotein (P) gene encodes four proteins. Three of these—P, V, and W—possess a common N-terminal domain but distinct C termini. These proteins interact with immune modulators. Previous studies demonstrated that P, V, and W bind STAT1 and STAT4 and that V also interacts with STAT2 but not with STAT3. The STAT1 and STAT2 interactions block interferon (IFN)-induced STAT tyrosine phosphorylation. To more fully characterize the interactions of P, V, and W with the STATs, we screened for interaction of each viral protein with STATs 1 to 6 by coimmunoprecipitation. We demonstrate that NiV P, V, and W interact with STAT4 through their common N-terminal domain and block STAT4 activity, based on a STAT4 response element reporter assay. Although none of the NiV proteins interact with STAT3 or STAT6, NiV V, but not P or W, interacts with STAT5 through its unique C terminus. Furthermore, the interaction of NiV V with STAT5 was not disrupted by overexpression of the N-terminal binding STAT1 or the C-terminal binding MDA5. NiV V also inhibits a STAT5 response element reporter assay. Residues 114 to 140 of the common N-terminal domain of the NiV P gene products were found to be sufficient to bind STAT1 and STAT4. Analysis of STAT1-STAT3 chimeras suggests that the P gene products target the STAT1 SH2 domain. When fused to GST, the 114-140 peptide is sufficient to decrease STAT1 phosphorylation in IFN-β-stimulated cells, suggesting that this peptide could potentially be fused to heterologous proteins to confer inhibition of STAT1- and STAT4-dependent responses. IMPORTANCE How Nipah virus (NiV) antagonizes innate immune responses is incompletely understood. The P gene of NiV encodes the P, V, and W proteins. These proteins have a common N-terminal sequence that is sufficient to bind to STAT1 and STAT2 and block IFN-induced signal transduction. This study sought to more fully understand how P, V, and W engage with the STAT family of transcription factors to influence their functions. The results identify a novel interaction of V with STAT5 and demonstrate V inhibition of STAT5 function. We also demonstrate that the common N-terminal residues 114 to 140 of P, V, and W are critical for inhibition of STAT1 and STAT4 function, map the interaction to the SH2 region of STAT1, and show that a fusion construct with this peptide significantly inhibits cytokine-induced STAT1 phosphorylation. These data clarify how these important virulence factors modulate innate antiviral defenses.https://journals.asm.org/doi/10.1128/mSphere.00449-20Nipah virusSTAT transcription factorsinterferon |
| spellingShingle | Timothy R. Keiffer Michael J. Ciancanelli Megan R. Edwards Christopher F. Basler Interactions of the Nipah Virus P, V, and W Proteins across the STAT Family of Transcription Factors mSphere Nipah virus STAT transcription factors interferon |
| title | Interactions of the Nipah Virus P, V, and W Proteins across the STAT Family of Transcription Factors |
| title_full | Interactions of the Nipah Virus P, V, and W Proteins across the STAT Family of Transcription Factors |
| title_fullStr | Interactions of the Nipah Virus P, V, and W Proteins across the STAT Family of Transcription Factors |
| title_full_unstemmed | Interactions of the Nipah Virus P, V, and W Proteins across the STAT Family of Transcription Factors |
| title_short | Interactions of the Nipah Virus P, V, and W Proteins across the STAT Family of Transcription Factors |
| title_sort | interactions of the nipah virus p v and w proteins across the stat family of transcription factors |
| topic | Nipah virus STAT transcription factors interferon |
| url | https://journals.asm.org/doi/10.1128/mSphere.00449-20 |
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