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| [PMID]: | 28768857 |
| [Au] Autor: | Nakagawa K; Kobayashi Y; Ito N; Suzuki Y; Okada K; Makino M; Goto H; Takahashi T; Sugiyama M |
| [Ad] Endereço: | The United Graduate School of Veterinary Sciences, Gifu University, Gifu, Japan. |
| [Ti] Título: | Molecular Function Analysis of Rabies Virus RNA Polymerase L Protein by Using an L Gene-Deficient Virus. |
| [So] Source: | J Virol;91(20), 2017 Oct 15. | | [Is] ISSN: | 1098-5514 |
| [Cp] País de publicação: | United States |
| [La] Idioma: | eng |
| [Ab] Resumo: | While the RNA-dependent RNA polymerase L protein of rabies virus (RABV), a member of the genus of the family , has potential to be a therapeutic target for rabies, the molecular functions of this protein have remained largely unknown. In this study, to obtain a novel experimental tool for molecular function analysis of the RABV L protein, we established by using a reverse genetics approach an L gene-deficient RABV (Nishi-ΔL/Nluc), which infects, propagates, and correspondingly produces NanoLuc luciferase in cultured neuroblastoma cells transfected to express the L protein. -Complementation with wild-type L protein, but not that with a functionally defective L protein mutant, efficiently supported luciferase production by Nishi-ΔL/Nluc, confirming its potential for function analysis of the L protein. Based on the findings obtained from comprehensive genetic analyses of L genes from various RABV and other lyssavirus species, we examined the functional importance of a highly conserved L protein region at positions 1914 to 1933 by a -complementation assay with Nishi-ΔL/Nluc and a series of L protein mutants. The results revealed that the amino acid sequence at positions 1929 to 1933 (NPYNE) is functionally important, and this was supported by other findings that this sequence is critical for binding of the L protein with its essential cofactor, P protein, and thus also for L protein's RNA polymerase activity. Our findings provide useful information for the development of an anti-RABV drug targeting the L-P protein interaction. To the best of our knowledge, this is the first report on the establishment of an L gene-deficient, reporter gene-expressing virus in all species of the order , also highlighting its applicability to a -complementation assay, which is useful for molecular function analyses of their L proteins. Moreover, this study revealed for the first time that the NPYNE sequence at positions 1929 to 1933 in the RABV L protein is important for L protein's interaction with the P protein, consistent with and extending the results of a previous study showing that the P protein-binding domain in the L protein is located in its C-terminal region, at positions 1562 to 2127. This study indicates that the NPYNE sequence is a promising target for the development of an inhibitor of viral RNA synthesis, which has high potential as a therapeutic drug for rabies. |
| [Mh] Termos MeSH primário: |
RNA Polimerases Dirigidas por DNA/genética
RNA Polimerases Dirigidas por DNA/metabolismo
Genes Virais
Vírus da Raiva/enzimologia
Proteínas Virais/genética
Proteínas Virais/metabolismo
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| [Mh] Termos MeSH secundário: |
Animais
Linhagem Celular
RNA Polimerases Dirigidas por DNA/química
Teste de Complementação Genética
Luciferases/biossíntese
Luciferases/genética
Lyssavirus/genética
Mutação
Fosfoproteínas/metabolismo
RNA Viral/genética
Vírus da Raiva/genética
Genética Reversa
Rhabdoviridae/genética
Proteínas Virais/química
Replicação Viral
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| [Pt] Tipo de publicação: | JOURNAL ARTICLE |
[Nm] Nome de substância:
| 0 (Phosphoproteins); 0 (RNA, Viral); 0 (Viral Proteins); EC 1.13.12.- (Luciferases); EC 2.7.7.48 (L protein, Rabies virus); EC 2.7.7.6 (DNA-Directed RNA Polymerases) |
| [Em] Mês de entrada: | 1710 |
| [Cu] Atualização por classe: | 171024 |
[Lr] Data última revisão:
| 171024 |
| [Sb] Subgrupo de revista: | IM |
| [Da] Data de entrada para processamento: | 170804 |
| [St] Status: | MEDLINE |
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