Mutation of CD2AP and SH3KBP1 Binding Motif in Alphavirus nsP3 Hypervariable Domain Results in Attenuated Virus
Infection by Chikungunya virus (CHIKV) of the Old World alphaviruses (family Togaviridae) in humans can cause arthritis and arthralgia. The virus encodes four non-structural proteins (nsP) (nsP1, nsp2, nsP3 and nsP4) that act as subunits of the virus replicase. These proteins also interact with nume...
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| Format: | Article |
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MDPI AG
2018-04-01
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| Series: | Viruses |
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| Online Access: | http://www.mdpi.com/1999-4915/10/5/226 |
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| author | Margit Mutso Ainhoa Moliner Morro Cecilia Smedberg Sergo Kasvandik Muriel Aquilimeba Mona Teppor Liisi Tarve Aleksei Lulla Valeria Lulla Sirle Saul Bastian Thaa Gerald M McInerney Andres Merits Margus Varjak |
| author_facet | Margit Mutso Ainhoa Moliner Morro Cecilia Smedberg Sergo Kasvandik Muriel Aquilimeba Mona Teppor Liisi Tarve Aleksei Lulla Valeria Lulla Sirle Saul Bastian Thaa Gerald M McInerney Andres Merits Margus Varjak |
| author_sort | Margit Mutso |
| collection | DOAJ |
| description | Infection by Chikungunya virus (CHIKV) of the Old World alphaviruses (family Togaviridae) in humans can cause arthritis and arthralgia. The virus encodes four non-structural proteins (nsP) (nsP1, nsp2, nsP3 and nsP4) that act as subunits of the virus replicase. These proteins also interact with numerous host proteins and some crucial interactions are mediated by the unstructured C-terminal hypervariable domain (HVD) of nsP3. In this study, a human cell line expressing EGFP tagged with CHIKV nsP3 HVD was established. Using quantitative proteomics, it was found that CHIKV nsP3 HVD can bind cytoskeletal proteins, including CD2AP, SH3KBP1, CAPZA1, CAPZA2 and CAPZB. The interaction with CD2AP was found to be most evident; its binding site was mapped to the second SH3 ligand-like element in nsP3 HVD. Further assessment indicated that CD2AP can bind to nsP3 HVDs of many different New and Old World alphaviruses. Mutation of the short binding element hampered the ability of the virus to establish infection. The mutation also abolished ability of CD2AP to co-localise with nsP3 and replication complexes of CHIKV; the same was observed for Semliki Forest virus (SFV) harbouring a similar mutation. Similar to CD2AP, its homolog SH3KBP1 also bound the identified motif in CHIKV and SFV nsP3. |
| first_indexed | 2024-04-13T02:31:02Z |
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| id | doaj.art-436e6e42498f4d28a75d5d8db1b0d5fb |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-04-13T02:31:02Z |
| publishDate | 2018-04-01 |
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| record_format | Article |
| series | Viruses |
| spelling | doaj.art-436e6e42498f4d28a75d5d8db1b0d5fb2022-12-22T03:06:35ZengMDPI AGViruses1999-49152018-04-0110522610.3390/v10050226v10050226Mutation of CD2AP and SH3KBP1 Binding Motif in Alphavirus nsP3 Hypervariable Domain Results in Attenuated VirusMargit Mutso0Ainhoa Moliner Morro1Cecilia Smedberg2Sergo Kasvandik3Muriel Aquilimeba4Mona Teppor5Liisi Tarve6Aleksei Lulla7Valeria Lulla8Sirle Saul9Bastian Thaa10Gerald M McInerney11Andres Merits12Margus Varjak13Institute of Technology, University of Tartu, 50411 Tartu, EstoniaDepartment of Microbiology, Tumor and Cell Biology, Karolinska Institutet, SE-171 77 Stockholm, SwedenDepartment of Microbiology, Tumor and Cell Biology, Karolinska Institutet, SE-171 77 Stockholm, SwedenInstitute of Technology, University of Tartu, 50411 Tartu, EstoniaInstitute of Technology, University of Tartu, 50411 Tartu, EstoniaInstitute of Technology, University of Tartu, 50411 Tartu, EstoniaInstitute of Technology, University of Tartu, 50411 Tartu, EstoniaInstitute of Technology, University of Tartu, 50411 Tartu, EstoniaInstitute of Technology, University of Tartu, 50411 Tartu, EstoniaInstitute of Technology, University of Tartu, 50411 Tartu, EstoniaDepartment of Microbiology, Tumor and Cell Biology, Karolinska Institutet, SE-171 77 Stockholm, SwedenDepartment of Microbiology, Tumor and Cell Biology, Karolinska Institutet, SE-171 77 Stockholm, SwedenInstitute of Technology, University of Tartu, 50411 Tartu, EstoniaInstitute of Technology, University of Tartu, 50411 Tartu, EstoniaInfection by Chikungunya virus (CHIKV) of the Old World alphaviruses (family Togaviridae) in humans can cause arthritis and arthralgia. The virus encodes four non-structural proteins (nsP) (nsP1, nsp2, nsP3 and nsP4) that act as subunits of the virus replicase. These proteins also interact with numerous host proteins and some crucial interactions are mediated by the unstructured C-terminal hypervariable domain (HVD) of nsP3. In this study, a human cell line expressing EGFP tagged with CHIKV nsP3 HVD was established. Using quantitative proteomics, it was found that CHIKV nsP3 HVD can bind cytoskeletal proteins, including CD2AP, SH3KBP1, CAPZA1, CAPZA2 and CAPZB. The interaction with CD2AP was found to be most evident; its binding site was mapped to the second SH3 ligand-like element in nsP3 HVD. Further assessment indicated that CD2AP can bind to nsP3 HVDs of many different New and Old World alphaviruses. Mutation of the short binding element hampered the ability of the virus to establish infection. The mutation also abolished ability of CD2AP to co-localise with nsP3 and replication complexes of CHIKV; the same was observed for Semliki Forest virus (SFV) harbouring a similar mutation. Similar to CD2AP, its homolog SH3KBP1 also bound the identified motif in CHIKV and SFV nsP3.http://www.mdpi.com/1999-4915/10/5/226alphavirusChikungunya virusCD2APSH3KBP1SH3 ligandproteomics |
| spellingShingle | Margit Mutso Ainhoa Moliner Morro Cecilia Smedberg Sergo Kasvandik Muriel Aquilimeba Mona Teppor Liisi Tarve Aleksei Lulla Valeria Lulla Sirle Saul Bastian Thaa Gerald M McInerney Andres Merits Margus Varjak Mutation of CD2AP and SH3KBP1 Binding Motif in Alphavirus nsP3 Hypervariable Domain Results in Attenuated Virus Viruses alphavirus Chikungunya virus CD2AP SH3KBP1 SH3 ligand proteomics |
| title | Mutation of CD2AP and SH3KBP1 Binding Motif in Alphavirus nsP3 Hypervariable Domain Results in Attenuated Virus |
| title_full | Mutation of CD2AP and SH3KBP1 Binding Motif in Alphavirus nsP3 Hypervariable Domain Results in Attenuated Virus |
| title_fullStr | Mutation of CD2AP and SH3KBP1 Binding Motif in Alphavirus nsP3 Hypervariable Domain Results in Attenuated Virus |
| title_full_unstemmed | Mutation of CD2AP and SH3KBP1 Binding Motif in Alphavirus nsP3 Hypervariable Domain Results in Attenuated Virus |
| title_short | Mutation of CD2AP and SH3KBP1 Binding Motif in Alphavirus nsP3 Hypervariable Domain Results in Attenuated Virus |
| title_sort | mutation of cd2ap and sh3kbp1 binding motif in alphavirus nsp3 hypervariable domain results in attenuated virus |
| topic | alphavirus Chikungunya virus CD2AP SH3KBP1 SH3 ligand proteomics |
| url | http://www.mdpi.com/1999-4915/10/5/226 |
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