Expression of the Rubella Virus Nonstructural Protein ORF and Demonstration of Proteolytic Processing
| Autor: | Lee D. Marr, Chin-Yen Wang, Teryl K. Frey |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
viruses
medicine.medical_treatment DNA Mutational Analysis Molecular Sequence Data Vaccinia virus Viral Nonstructural Proteins Biology Open Reading Frames Cricetinae Virology Gene expression medicine Animals T7 RNA polymerase Cloning Molecular Cells Cultured Sequence Deletion NS3 Protease Expression vector Base Sequence Sequence Homology Amino Acid Molecular biology Cysteine protease Recombinant Proteins NS2-3 protease Cysteine Endopeptidases Open reading frame DNA Viral Mutagenesis Site-Directed Protein Processing Post-Translational Rubella virus medicine.drug |
| Zdroj: | Virology. 198:586-592 |
| ISSN: | 0042-6822 |
| DOI: | 10.1006/viro.1994.1070 |
| Popis: | To analyze the proteins produced from the rubella virus (RUB) nonstructural protein open reading frame (NSP-ORF), a DNA containing the RUB NSP-ORF was introduced into the expression vector pTM3 in which the sequences to be expressed are downstream from a T7 RNA polymerase promoter. In cells infected with a vaccinia virus recombinant which expresses T7 RNA polymerase and transfected with this plasmid, three RUB-specific products with electrophoretic mobilities of 200, 150, and 97 kDa were clearly visible. By computer alignment, the presence of a cysteine protease was predicted within the NSP-ORF (A. E. Gorbalenya et al., FEBS Lett. 288, 201-205, 1991). When the Cys proposed as the catalytic residue of this protease (Cys1151) was mutated to a Gly, only the 200-kDa product was produced, demonstrating that the Cys is important in the activity of the protease responsible for the processing of the RUB NSPs and that the 150- and 97-kDa species are processing products. Transfections with deletion mutants revealed that the 150-kDa processing product is derived from the amino-terminal two-thirds of the ORF and that both the protease and the cleavage site on the COOH-terminal side of the 150-kDa product are between amino acids 1005 and 1507 of the ORF. |
| Databáze: | OpenAIRE |
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