Mutagenesis of the yellow fever virus NS2B/3 cleavage site: determinants of cleavage site specificity and effects on polyprotein processing and viral replication
| ISSN: | 1098-5514 0022-538X |
|---|---|
| DOI: | 10.1128/jvi.69.3.1600-1605.1995 |
| Přístupová URL adresa: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::753ba01b14583018f60f0b340b761964 https://doi.org/10.1128/jvi.69.3.1600-1605.1995 |
| Rights: | OPEN |
| Přírůstkové číslo: | edsair.doi.dedup.....753ba01b14583018f60f0b340b761964 |
| Autor: | Charles M. Rice, A Nestorowicz, T J Chambers |
| Rok vydání: | 1995 |
| Předmět: |
Cleavage factor
viruses Molecular Sequence Data Immunology Cleavage and polyadenylation specificity factor Viral Nonstructural Proteins Virus Replication Cleavage (embryo) Microbiology Structure-Activity Relationship Viral Proteins Virology Amino Acid Sequence NS3 Cleavage stimulation factor biology Proteins biology.organism_classification Molecular biology NS2-3 protease Flavivirus Viral replication Insect Science Mutagenesis Site-Directed Yellow fever virus Protein Processing Post-Translational Research Article |
| Zdroj: | Journal of Virology. 69:1600-1605 |
| ISSN: | 1098-5514 0022-538X |
| DOI: | 10.1128/jvi.69.3.1600-1605.1995 |
| Popis: | The determinants of cleavage site specificity of the yellow fever virus (YF) NS3 proteinase for its 2B/3 cleavage site have been studied by using site-directed mutagenesis. Mutations at residues within the GARR decreases S sequence were tested for effects on cis cleavage of an NS2B-3(181) polyprotein during cell-free translation. At the P1 position, only the conservative substitution R-->K exhibited significant levels of cleavage. Conservative and nonconservative substitutions were tolerated at the P1' and P2 positions, resulting in intermediate levels of cleavage. Substitutions at the P3 and P4 positions had no effects on cleavage efficiency in the cell-free assay. Processing at other dibasic sites was studied by using transient expression of a sig2A-5(356) polyprotein. Cleavage at the 2B/3 site was not required for processing at downstream sites. However, increased accumulation of high-molecular-weight viral polyproteins was generally observed for mutations which reduced cleavage efficiency at the 2B/3 site. Several mutations were also tested for their effects on viral replication. Virus was not recovered from substitutions which blocked or substantially reduced cleavage in the cell-free assay, suggesting that efficient cleavage at the 2B/3 site is required for flavivirus replication. |
| Databáze: | OpenAIRE |
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